Does human CD16 bind to rat anti-mouse IgG1? Specifically, will human NK cells mediate ADCC against targets labelled with rat anti-mouse IgG1? Thanks, Michael A. Sheard
Posts in Antibody Discussion Forum
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Thanks for the responses. Seems there are limited options with a fusion partner that results in unstable hybridomas, or challenge of making a phage library. Looks like another group made improvements to Knightβs myeloma worth looking into. Jason M. Goldstein Founder 7560 Portbury Park Lane Suwanee, GA 30024 415-519-4493 <email obscured> | www.annularbio.com
> On Oct 30, 2018, at 5:10 PM, Mark Sleeman <email obscured>> wrote: > > Hi > This was a timely post. We have also had lots of requests and are > starting some work in this area. We will seek out those lines > mentioned and keep people informed of progress. any additional > information would be greatly appreciated > Thanks > Mark > > > > Sent from my iPhone > >> On 31 Oct 2018, at 06:15, David Blum <email obscured>> wrote: >> >> The 240E line is available from Katherine Knight at Loyola University via a >> license. We have plans to use it, but have been told that it results in >> unstable hybridomas. It is no longer available from the ATCC, BTW. Also, >> I'm not a lawyer, but I believe we are restricted to our own use at our own >> institution. >> >> https://en.wikipedia.org/wiki/Rabbit_hybridoma >> >> >> >> On Tue, Oct 30, 2018 at 3:51 PM Jason Goldstein <email obscured>> >> wrote: >> >>> >>> Does anyone have experience with development of rabbit monoclonals? I am >>> unable to identify a myeloma line and unsure if still under license >>> agreement. Can rabbit B-cells be fused by electrofusion similar to murine? >>> Thanks, >>> Jason >>> >>> >>> >>> Jason M. Goldstein >>> Founder >>> >>> 7560 Portbury Park Lane >>> Suwanee, GA 30024 >>> >>> 415-519-4493 >>> >>> <email obscured> | www.annularbio.com >>> >>> >>>> On Oct 12, 2018, at 4:47 PM, Weis-Garcia, Frances/Sloan Kettering >>> Institute <email obscured>> wrote: >>>> >>>> FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson >>> ImmunoReagents ... and they definitely have an anti-human, so we are good. >>>> >>>> ο»Ώ-----Original Message----- >>>> From: Antibody Discussion Forum <email obscured>> on behalf of >>> Laura Bover <email obscured>> >>>> Reply-To: <email obscured>> >>>> Date: Friday, October 12, 2018 at 4:37 PM >>>> To: <email obscured>" <email obscured>> >>>> Cc: Lloyd Bourne <email obscured>> >>>> Subject: Re: [ABRF Ab Forum] ClonePix >>>> >>>> Hi Philip, >>>> We also have a Clonepix 2 but so far only used for hybridomas. >>>> You will need the Detection fluorescence reagent from Molecular >>> Devices that will help in the selection and pick up of the human IgG good >>> producer clones. And I am not sure that they have a detection reagent for >>> human IgGs. We use the murine detection reagent only. >>>> >>>> You may also try old fashion cloning by limited dilution. And test by >>> ELISA and/or FACS the supernatants to select the best producers. We do a >>> titration with supernatants and pick the bests. >>>> I hope this helps. >>>> >>>> Thanks! >>>> >>>> >>>> Laura Bover, PhD >>>> Professor >>>> Director Monoclonal Antibody Core Facility >>>> University of Texas M.D.Anderson Cancer Center >>>> Associate member of The University of Texas Graduate School of >>> Biomedical Sciences at Houston >>>> Immunology Department/ Genomics Medicine Department >>>> 7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039 >>>> Houston- TX 77054 >>>> T 713-563-2281/ 713-563-3301 >>>> F 713-563-3276 >>>> <email obscured> >>> <email obscured> >>>> >>>> >>>> >>>> On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of >>> Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> >>> on behalf of <email obscured>> wrote: >>>> >>>> Everyone in this discussion forum - Anyone have experience >>> growing CHO cells in SS media? >>>> >>>> Philip, While you have them if you want to play around with >>> culturing CHO cells in SS media that would save time. Molecular Devices >>> and Stem Cell Technologies should have something ... maybe Fisher too. We >>> have not, so we will need to run some pilots. >>>> >>>> Frances >>>> >>>> -----Original Message----- >>>> From: Antibody Discussion Forum <email obscured>> on >>> behalf of Phillip Gao <email obscured>> >>>> Reply-To: <email obscured>> >>>> Date: Friday, October 12, 2018 at 2:08 PM >>>> To: <email obscured>" <email obscured>> >>>> Cc: Lloyd Bourne <email obscured>> >>>> Subject: Re: [ABRF Ab Forum] ClonePix >>>> >>>> Hi, Frances: >>>> >>>> Thank you so much! >>>> >>>> Yes, the antibody is humanized. >>>> >>>> I need the PI deliver the cells and medium to me, then make a >>> bank. I will also take a close look of the manual and related materials so >>> that know what I am doing before I send you everything. I will defiantly >>> test the mycoplasma before I send them to you. >>>> >>>> Have a nice weekend. >>>> >>>> Philip >>>> >>>> -----Original Message----- >>>> From: Antibody Discussion Forum <email obscured>> On >>> Behalf Of Weis-Garcia, Frances/Sloan Kettering Institute >>>> Sent: Friday, October 12, 2018 12:55 PM >>>> To: <email obscured> >>>> Cc: Lloyd Bourne <email obscured>> >>>> Subject: Re: [ABRF Ab Forum] ClonePix >>>> >>>> We have one, and are open to trying. >>>> We have only grown hybridomas but if the CHO are in >>> suspension it should be fine. >>>> >>>> If you send the media (liquid and semisolid) and cells, we >>> can give it try and only charge you if it works. Fair? >>>> >>>> Have the cells been tested for mycoplasma? >>>> Please confirm the antibody has a human Fc domain. >>>> If you do not tell me the antigen, I think we can avoid >>> signing any MTA. >>>> >>>> Frances >>>> >>>> -----Original Message----- >>>> From: Antibody Discussion Forum <email obscured>> on >>> behalf of Phillip Gao <email obscured>> >>>> Reply-To: <email obscured>> >>>> Date: Friday, October 12, 2018 at 1:10 PM >>>> To: <email obscured>" <email obscured>> >>>> Subject: [ABRF Ab Forum] ClonePix >>>> >>>> Dear Friends: >>>> >>>> I have a pool of mixed antibody producing CHO suspension >>> cells. The medium is commercially available. But the problem is the yield >>> of the antibody (IgG) is very low, <100ug/l. We have the purified antigen. >>>> >>>> Does anyone have a ClonePix and how much would it cost >>> for the service? Or any other better approaches? >>>> >>>> Rob at Vanderbilt had one and it is no longer in service. >>>> >>>> Thank you for your help. >>>> >>>> Philip >>>> >>>> >>>> Philip Gao, Ph. D. >>>> Director, COBRE Protein Production Group >>>> Del Shankel Structural Biology Center >>>> 2034 Becker Drive, Room 1095 >>>> University of Kansas >>>> Lawrence, KS 66047 >>>> Phone: (785) 864-1825 (office); (785) 864-3406 (lab) >>>> Fax: (785) 864-8141 >>>> Website: http://psf.cobre.ku.edu/cores/ppg/about >>>> >>>> >>>> >>>> ββ >>>> View topic >>> http://list.abrf.org/r/topic/3eLY0BxEOzs635Ju6KtFK9 >>>> Leave group <email obscured> >>> ?Subject=Unsubscribe >>>> >>>> >>>> >>>> >>>> >>>> ββ >>>> View topic >>> http://list.abrf.org/r/topic/4Cl1dkiqYfV569lP4gjVwi >>>> Leave group <email obscured>?Subject=Unsubscribe >>>> >>>> >>>> ββ >>>> View topic >>> http://list.abrf.org/r/topic/5iJHbs5g1ZT3bM1RD8B1SZ >>>> Leave group <email obscured>?Subject=Unsubscribe >>>> >>>> >>>> >>>> >>>> >>>> ββ >>>> View topic http://list.abrf.org/r/topic/dUj44DBjpYNqPcUFvi7zG >>>> Leave group <email obscured>?Subject=Unsubscribe >>>> >>>> >>>> >>>> The information contained in this e-mail message may be privileged, >>> confidential, and/or protected from disclosure. This e-mail message may >>> contain protected health information (PHI); dissemination of PHI should >>> comply with applicable federal and state laws. If you are not the intended >>> recipient, or an authorized representative of the intended recipient, any >>> further review, disclosure, use, dissemination, distribution, or copying of >>> this message or any attachment (or the information contained therein) is >>> strictly prohibited. If you think that you have received this e-mail >>> message in error, please notify the sender by return e-mail and delete all >>> references to it and its contents from your systems. >>>> >>>> ββ >>>> View topic http://list.abrf.org/r/topic/2nBJWp9ehr0eVk9quw6EmC >>>> Leave group <email obscured>?Subject=Unsubscribe >>>> >>>> >>>> >>>> >>>> >>>> ββ >>>> View topic http://list.abrf.org/r/topic/3g3fsRz2INNio4DTv6nVle >>>> Leave group <email obscured>?Subject=Unsubscribe >>>> >>> >>> ββ >>> View topic http://list.abrf.org/r/topic/FImBfKy04YbD7SUwvCbaV >>> Leave group <email obscured>?Subject=Unsubscribe >>> >>> >> >> ββ >> View topic http://list.abrf.org/r/topic/7yCmsjYHNvb0QafxEBWQVd >> Leave group <email obscured>?Subject=Unsubscribe >> > > ββ > View topic http://list.abrf.org/r/topic/7tAJuIvyudLfNA9NTYZ0wK > Leave group <email obscured>?Subject=Unsubscribe >
Hi This was a timely post. We have also had lots of requests and are starting some work in this area. We will seek out those lines mentioned and keep people informed of progress. any additional information would be greatly appreciated Thanks Mark Sent from my iPhone
> On 31 Oct 2018, at 06:15, David Blum <email obscured>> wrote: > > The 240E line is available from Katherine Knight at Loyola University via a > license. We have plans to use it, but have been told that it results in > unstable hybridomas. It is no longer available from the ATCC, BTW. Also, > I'm not a lawyer, but I believe we are restricted to our own use at our own > institution. > > https://en.wikipedia.org/wiki/Rabbit_hybridoma > > > > On Tue, Oct 30, 2018 at 3:51 PM Jason Goldstein <email obscured>> > wrote: > >> >> Does anyone have experience with development of rabbit monoclonals? I am >> unable to identify a myeloma line and unsure if still under license >> agreement. Can rabbit B-cells be fused by electrofusion similar to murine? >> Thanks, >> Jason >> >> >> >> Jason M. Goldstein >> Founder >> >> 7560 Portbury Park Lane >> Suwanee, GA 30024 >> >> 415-519-4493 >> >> <email obscured> | www.annularbio.com >> >> >>> On Oct 12, 2018, at 4:47 PM, Weis-Garcia, Frances/Sloan Kettering >> Institute <email obscured>> wrote: >>> >>> FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson >> ImmunoReagents ... and they definitely have an anti-human, so we are good. >>> >>> ο»Ώ-----Original Message----- >>> From: Antibody Discussion Forum <email obscured>> on behalf of >> Laura Bover <email obscured>> >>> Reply-To: <email obscured>> >>> Date: Friday, October 12, 2018 at 4:37 PM >>> To: <email obscured>" <email obscured>> >>> Cc: Lloyd Bourne <email obscured>> >>> Subject: Re: [ABRF Ab Forum] ClonePix >>> >>> Hi Philip, >>> We also have a Clonepix 2 but so far only used for hybridomas. >>> You will need the Detection fluorescence reagent from Molecular >> Devices that will help in the selection and pick up of the human IgG good >> producer clones. And I am not sure that they have a detection reagent for >> human IgGs. We use the murine detection reagent only. >>> >>> You may also try old fashion cloning by limited dilution. And test by >> ELISA and/or FACS the supernatants to select the best producers. We do a >> titration with supernatants and pick the bests. >>> I hope this helps. >>> >>> Thanks! >>> >>> >>> Laura Bover, PhD >>> Professor >>> Director Monoclonal Antibody Core Facility >>> University of Texas M.D.Anderson Cancer Center >>> Associate member of The University of Texas Graduate School of >> Biomedical Sciences at Houston >>> Immunology Department/ Genomics Medicine Department >>> 7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039 >>> Houston- TX 77054 >>> T 713-563-2281/ 713-563-3301 >>> F 713-563-3276 >>> <email obscured> >> <email obscured> >>> >>> >>> >>> On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of >> Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> >> on behalf of <email obscured>> wrote: >>> >>> Everyone in this discussion forum - Anyone have experience >> growing CHO cells in SS media? >>> >>> Philip, While you have them if you want to play around with >> culturing CHO cells in SS media that would save time. Molecular Devices >> and Stem Cell Technologies should have something ... maybe Fisher too. We >> have not, so we will need to run some pilots. >>> >>> Frances >>> >>> -----Original Message----- >>> From: Antibody Discussion Forum <email obscured>> on >> behalf of Phillip Gao <email obscured>> >>> Reply-To: <email obscured>> >>> Date: Friday, October 12, 2018 at 2:08 PM >>> To: <email obscured>" <email obscured>> >>> Cc: Lloyd Bourne <email obscured>> >>> Subject: Re: [ABRF Ab Forum] ClonePix >>> >>> Hi, Frances: >>> >>> Thank you so much! >>> >>> Yes, the antibody is humanized. >>> >>> I need the PI deliver the cells and medium to me, then make a >> bank. I will also take a close look of the manual and related materials so >> that know what I am doing before I send you everything. I will defiantly >> test the mycoplasma before I send them to you. >>> >>> Have a nice weekend. >>> >>> Philip >>> >>> -----Original Message----- >>> From: Antibody Discussion Forum <email obscured>> On >> Behalf Of Weis-Garcia, Frances/Sloan Kettering Institute >>> Sent: Friday, October 12, 2018 12:55 PM >>> To: <email obscured> >>> Cc: Lloyd Bourne <email obscured>> >>> Subject: Re: [ABRF Ab Forum] ClonePix >>> >>> We have one, and are open to trying. >>> We have only grown hybridomas but if the CHO are in >> suspension it should be fine. >>> >>> If you send the media (liquid and semisolid) and cells, we >> can give it try and only charge you if it works. Fair? >>> >>> Have the cells been tested for mycoplasma? >>> Please confirm the antibody has a human Fc domain. >>> If you do not tell me the antigen, I think we can avoid >> signing any MTA. >>> >>> Frances >>> >>> -----Original Message----- >>> From: Antibody Discussion Forum <email obscured>> on >> behalf of Phillip Gao <email obscured>> >>> Reply-To: <email obscured>> >>> Date: Friday, October 12, 2018 at 1:10 PM >>> To: <email obscured>" <email obscured>> >>> Subject: [ABRF Ab Forum] ClonePix >>> >>> Dear Friends: >>> >>> I have a pool of mixed antibody producing CHO suspension >> cells. The medium is commercially available. But the problem is the yield >> of the antibody (IgG) is very low, <100ug/l. We have the purified antigen. >>> >>> Does anyone have a ClonePix and how much would it cost >> for the service? Or any other better approaches? >>> >>> Rob at Vanderbilt had one and it is no longer in service. >>> >>> Thank you for your help. >>> >>> Philip >>> >>> >>> Philip Gao, Ph. D. >>> Director, COBRE Protein Production Group >>> Del Shankel Structural Biology Center >>> 2034 Becker Drive, Room 1095 >>> University of Kansas >>> Lawrence, KS 66047 >>> Phone: (785) 864-1825 (office); (785) 864-3406 (lab) >>> Fax: (785) 864-8141 >>> Website: http://psf.cobre.ku.edu/cores/ppg/about >>> >>> >>> >>> ββ >>> View topic >> http://list.abrf.org/r/topic/3eLY0BxEOzs635Ju6KtFK9 >>> Leave group <email obscured> >> ?Subject=Unsubscribe >>> >>> >>> >>> >>> >>> ββ >>> View topic >> http://list.abrf.org/r/topic/4Cl1dkiqYfV569lP4gjVwi >>> Leave group <email obscured>?Subject=Unsubscribe >>> >>> >>> ββ >>> View topic >> http://list.abrf.org/r/topic/5iJHbs5g1ZT3bM1RD8B1SZ >>> Leave group <email obscured>?Subject=Unsubscribe >>> >>> >>> >>> >>> >>> ββ >>> View topic http://list.abrf.org/r/topic/dUj44DBjpYNqPcUFvi7zG >>> Leave group <email obscured>?Subject=Unsubscribe >>> >>> >>> >>> The information contained in this e-mail message may be privileged, >> confidential, and/or protected from disclosure. This e-mail message may >> contain protected health information (PHI); dissemination of PHI should >> comply with applicable federal and state laws. If you are not the intended >> recipient, or an authorized representative of the intended recipient, any >> further review, disclosure, use, dissemination, distribution, or copying of >> this message or any attachment (or the information contained therein) is >> strictly prohibited. If you think that you have received this e-mail >> message in error, please notify the sender by return e-mail and delete all >> references to it and its contents from your systems. >>> >>> ββ >>> View topic http://list.abrf.org/r/topic/2nBJWp9ehr0eVk9quw6EmC >>> Leave group <email obscured>?Subject=Unsubscribe >>> >>> >>> >>> >>> >>> ββ >>> View topic http://list.abrf.org/r/topic/3g3fsRz2INNio4DTv6nVle >>> Leave group <email obscured>?Subject=Unsubscribe >>> >> >> ββ >> View topic http://list.abrf.org/r/topic/FImBfKy04YbD7SUwvCbaV >> Leave group <email obscured>?Subject=Unsubscribe >> >> > > ββ > View topic http://list.abrf.org/r/topic/7yCmsjYHNvb0QafxEBWQVd > Leave group <email obscured>?Subject=Unsubscribe >
The 240E line is available from Katherine Knight at Loyola University via a license. We have plans to use it, but have been told that it results in unstable hybridomas. It is no longer available from the ATCC, BTW. Also, I'm not a lawyer, but I believe we are restricted to our own use at our own institution. https://en.wikipedia.org/wiki/Rabbit_hybridoma On Tue, Oct 30, 2018 at 3:51 PM Jason Goldstein <email obscured>> wrote:
> > Does anyone have experience with development of rabbit monoclonals? I am > unable to identify a myeloma line and unsure if still under license > agreement. Can rabbit B-cells be fused by electrofusion similar to murine? > Thanks, > Jason > > > > Jason M. Goldstein > Founder > > 7560 Portbury Park Lane > Suwanee, GA 30024 > > 415-519-4493 > > <email obscured> | www.annularbio.com > > > > On Oct 12, 2018, at 4:47 PM, Weis-Garcia, Frances/Sloan Kettering > Institute <email obscured>> wrote: > > > > FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson > ImmunoReagents ... and they definitely have an anti-human, so we are good. > > > > ο»Ώ-----Original Message----- > > From: Antibody Discussion Forum <email obscured>> on behalf of > Laura Bover <email obscured>> > > Reply-To: <email obscured>> > > Date: Friday, October 12, 2018 at 4:37 PM > > To: <email obscured>" <email obscured>> > > Cc: Lloyd Bourne <email obscured>> > > Subject: Re: [ABRF Ab Forum] ClonePix > > > > Hi Philip, > > We also have a Clonepix 2 but so far only used for hybridomas. > > You will need the Detection fluorescence reagent from Molecular > Devices that will help in the selection and pick up of the human IgG good > producer clones. And I am not sure that they have a detection reagent for > human IgGs. We use the murine detection reagent only. > > > > You may also try old fashion cloning by limited dilution. And test by > ELISA and/or FACS the supernatants to select the best producers. We do a > titration with supernatants and pick the bests. > > I hope this helps. > > > > Thanks! > > > > > > Laura Bover, PhD > > Professor > > Director Monoclonal Antibody Core Facility > > University of Texas M.D.Anderson Cancer Center > > Associate member of The University of Texas Graduate School of > Biomedical Sciences at Houston > > Immunology Department/ Genomics Medicine Department > > 7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039 > > Houston- TX 77054 > > T 713-563-2281/ 713-563-3301 > > F 713-563-3276 > > <email obscured> > <email obscured> > > > > > > > > On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of > Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> > on behalf of <email obscured>> wrote: > > > > Everyone in this discussion forum - Anyone have experience > growing CHO cells in SS media? > > > > Philip, While you have them if you want to play around with > culturing CHO cells in SS media that would save time. Molecular Devices > and Stem Cell Technologies should have something ... maybe Fisher too. We > have not, so we will need to run some pilots. > > > > Frances > > > > -----Original Message----- > > From: Antibody Discussion Forum <email obscured>> on > behalf of Phillip Gao <email obscured>> > > Reply-To: <email obscured>> > > Date: Friday, October 12, 2018 at 2:08 PM > > To: <email obscured>" <email obscured>> > > Cc: Lloyd Bourne <email obscured>> > > Subject: Re: [ABRF Ab Forum] ClonePix > > > > Hi, Frances: > > > > Thank you so much! > > > > Yes, the antibody is humanized. > > > > I need the PI deliver the cells and medium to me, then make a > bank. I will also take a close look of the manual and related materials so > that know what I am doing before I send you everything. I will defiantly > test the mycoplasma before I send them to you. > > > > Have a nice weekend. > > > > Philip > > > > -----Original Message----- > > From: Antibody Discussion Forum <email obscured>> On > Behalf Of Weis-Garcia, Frances/Sloan Kettering Institute > > Sent: Friday, October 12, 2018 12:55 PM > > To: <email obscured> > > Cc: Lloyd Bourne <email obscured>> > > Subject: Re: [ABRF Ab Forum] ClonePix > > > > We have one, and are open to trying. > > We have only grown hybridomas but if the CHO are in > suspension it should be fine. > > > > If you send the media (liquid and semisolid) and cells, we > can give it try and only charge you if it works. Fair? > > > > Have the cells been tested for mycoplasma? > > Please confirm the antibody has a human Fc domain. > > If you do not tell me the antigen, I think we can avoid > signing any MTA. > > > > Frances > > > > -----Original Message----- > > From: Antibody Discussion Forum <email obscured>> on > behalf of Phillip Gao <email obscured>> > > Reply-To: <email obscured>> > > Date: Friday, October 12, 2018 at 1:10 PM > > To: <email obscured>" <email obscured>> > > Subject: [ABRF Ab Forum] ClonePix > > > > Dear Friends: > > > > I have a pool of mixed antibody producing CHO suspension > cells. The medium is commercially available. But the problem is the yield > of the antibody (IgG) is very low, <100ug/l. We have the purified antigen. > > > > Does anyone have a ClonePix and how much would it cost > for the service? Or any other better approaches? > > > > Rob at Vanderbilt had one and it is no longer in service. > > > > Thank you for your help. > > > > Philip > > > > > > Philip Gao, Ph. D. > > Director, COBRE Protein Production Group > > Del Shankel Structural Biology Center > > 2034 Becker Drive, Room 1095 > > University of Kansas > > Lawrence, KS 66047 > > Phone: (785) 864-1825 (office); (785) 864-3406 (lab) > > Fax: (785) 864-8141 > > Website: http://psf.cobre.ku.edu/cores/ppg/about > > > > > > > > ββ > > View topic > http://list.abrf.org/r/topic/3eLY0BxEOzs635Ju6KtFK9 > > Leave group <email obscured> > ?Subject=Unsubscribe > > > > > > > > > > > > ββ > > View topic > http://list.abrf.org/r/topic/4Cl1dkiqYfV569lP4gjVwi > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > > View topic > http://list.abrf.org/r/topic/5iJHbs5g1ZT3bM1RD8B1SZ > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > > > ββ > > View topic http://list.abrf.org/r/topic/dUj44DBjpYNqPcUFvi7zG > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > The information contained in this e-mail message may be privileged, > confidential, and/or protected from disclosure. This e-mail message may > contain protected health information (PHI); dissemination of PHI should > comply with applicable federal and state laws. If you are not the intended > recipient, or an authorized representative of the intended recipient, any > further review, disclosure, use, dissemination, distribution, or copying of > this message or any attachment (or the information contained therein) is > strictly prohibited. If you think that you have received this e-mail > message in error, please notify the sender by return e-mail and delete all > references to it and its contents from your systems. > > > > ββ > > View topic http://list.abrf.org/r/topic/2nBJWp9ehr0eVk9quw6EmC > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > > > ββ > > View topic http://list.abrf.org/r/topic/3g3fsRz2INNio4DTv6nVle > > Leave group <email obscured>?Subject=Unsubscribe > > > > ββ > View topic http://list.abrf.org/r/topic/FImBfKy04YbD7SUwvCbaV > Leave group <email obscured>?Subject=Unsubscribe > >
We tried and never had great success.
After a good bit of work we were able to get Christophe Rader's system up and
running. A specific breed of rabbit is immunized then bone marrow and spleen
is used to make a display library that is panned. Christophe is at Scripps
Florida.
Rob
ο»ΏOn 10/30/18, 2:52 PM, "Antibody Discussion Forum on behalf of Jason Goldstein"
<email obscured> on behalf of <email obscured>> wrote:
Does anyone have experience with development of rabbit monoclonals? I am
unable to identify a myeloma line and unsure if still under license agreement.
Can rabbit B-cells be fused by electrofusion similar to murine?
Thanks,
Jason
Jason M. Goldstein
Founder
7560 Portbury Park Lane
Suwanee, GA 30024
415-519-4493
<email obscured> |
https://na01.safelinks.protection.outlook.com/?url=www.annularbio.com&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cc16cbddfde244834ab5c08d63ea126bc%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636765259200041191&sdata=DGU9P%2BRQs7xMtM4AuUg%2Bp9JV56sSSJ%2B8ewPfwvc42Ao%3D&reserved=0
> On Oct 12, 2018, at 4:47 PM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
>
> FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson
ImmunoReagents ... and they definitely have an anti-human, so we are good.
>
> -----Original Message-----
> From: Antibody Discussion Forum <email obscured>> on behalf of
Laura Bover <email obscured>>
> Reply-To: <email obscured>>
> Date: Friday, October 12, 2018 at 4:37 PM
> To: <email obscured>" <email obscured>>
> Cc: Lloyd Bourne <email obscured>>
> Subject: Re: [ABRF Ab Forum] ClonePix
>
> Hi Philip,
> We also have a Clonepix 2 but so far only used for hybridomas.
> You will need the Detection fluorescence reagent from Molecular
Devices that will help in the selection and pick up of the human IgG good
producer clones. And I am not sure that they have a detection reagent for human
IgGs. We use the murine detection reagent only.
>
> You may also try old fashion cloning by limited dilution. And test by
ELISA and/or FACS the supernatants to select the best producers. We do a
titration with supernatants and pick the bests.
> I hope this helps.
>
> Thanks!
>
>
> Laura Bover, PhD
> Professor
> Director Monoclonal Antibody Core Facility
> University of Texas M.D.Anderson Cancer Center
> Associate member of The University of Texas Graduate School of
Biomedical Sciences at Houston
> Immunology Department/ Genomics Medicine Department
> 7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039
> Houston- TX 77054
> T 713-563-2281/ 713-563-3301
> F 713-563-3276
> <email obscured>
<email obscured>>
>
>
> On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of
Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> on
behalf of <email obscured>> wrote:
>
> Everyone in this discussion forum - Anyone have experience growing
CHO cells in SS media?
>
> Philip, While you have them if you want to play around with
culturing CHO cells in SS media that would save time. Molecular Devices and
Stem Cell Technologies should have something ... maybe Fisher too. We have
not, so we will need to run some pilots.
>
> Frances
>
> -----Original Message-----
> From: Antibody Discussion Forum <email obscured>> on behalf
of Phillip Gao <email obscured>>
> Reply-To: <email obscured>>
> Date: Friday, October 12, 2018 at 2:08 PM
> To: <email obscured>" <email obscured>>
> Cc: Lloyd Bourne <email obscured>>
> Subject: Re: [ABRF Ab Forum] ClonePix
>
> Hi, Frances:
>
> Thank you so much!
>
> Yes, the antibody is humanized.
>
> I need the PI deliver the cells and medium to me, then make a
bank. I will also take a close look of the manual and related materials so that
know what I am doing before I send you everything. I will defiantly test the
mycoplasma before I send them to you.
>
> Have a nice weekend.
>
> Philip
>
> -----Original Message-----
> From: Antibody Discussion Forum <email obscured>> On
Behalf Of Weis-Garcia, Frances/Sloan Kettering Institute
> Sent: Friday, October 12, 2018 12:55 PM
> To: <email obscured>
> Cc: Lloyd Bourne <email obscured>>
> Subject: Re: [ABRF Ab Forum] ClonePix
>
> We have one, and are open to trying.
> We have only grown hybridomas but if the CHO are in suspension
it should be fine.
>
> If you send the media (liquid and semisolid) and cells, we can
give it try and only charge you if it works. Fair?
>
> Have the cells been tested for mycoplasma?
> Please confirm the antibody has a human Fc domain.
> If you do not tell me the antigen, I think we can avoid
signing any MTA.
>
> Frances
>
> -----Original Message-----
> From: Antibody Discussion Forum <email obscured>> on
behalf of Phillip Gao <email obscured>>
> Reply-To: <email obscured>>
> Date: Friday, October 12, 2018 at 1:10 PM
> To: <email obscured>" <email obscured>>
> Subject: [ABRF Ab Forum] ClonePix
>
> Dear Friends:
>
> I have a pool of mixed antibody producing CHO suspension
cells. The medium is commercially available. But the problem is the yield of
the antibody (IgG) is very low, <100ug/l. We have the purified antigen.
>
> Does anyone have a ClonePix and how much would it cost for
the service? Or any other better approaches?
>
> Rob at Vanderbilt had one and it is no longer in service.
>
> Thank you for your help.
>
> Philip
>
>
> Philip Gao, Ph. D.
> Director, COBRE Protein Production Group
> Del Shankel Structural Biology Center
> 2034 Becker Drive, Room 1095
> University of Kansas
> Lawrence, KS 66047
> Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
> Fax: (785) 864-8141
> Website:
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fpsf.cobre.ku.edu%2Fcores%2Fppg%2Fabout&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cc16cbddfde244834ab5c08d63ea126bc%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636765259200041191&sdata=V6Q9dqtWwhcERhM7Rg83tu%2FVERL9KNCElof9feRnXRk%3D&reserved=0
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Does anyone have experience with development of rabbit monoclonals? I am unable to identify a myeloma line and unsure if still under license agreement. Can rabbit B-cells be fused by electrofusion similar to murine? Thanks, Jason Jason M. Goldstein Founder 7560 Portbury Park Lane Suwanee, GA 30024 415-519-4493 <email obscured> | www.annularbio.com > On Oct 12, 2018, at 4:47 PM, Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: > > FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson ImmunoReagents ... and they definitely have an anti-human, so we are good. > > ο»Ώ-----Original Message----- > From: Antibody Discussion Forum <email obscured>> on behalf of Laura Bover <email obscured>> > Reply-To: <email obscured>> > Date: Friday, October 12, 2018 at 4:37 PM > To: <email obscured>" <email obscured>> > Cc: Lloyd Bourne <email obscured>> > Subject: Re: [ABRF Ab Forum] ClonePix > > Hi Philip, > We also have a Clonepix 2 but so far only used for hybridomas. > You will need the Detection fluorescence reagent from Molecular Devices that will help in the selection and pick up of the human IgG good producer clones. And I am not sure that they have a detection reagent for human IgGs. We use the murine detection reagent only. > > You may also try old fashion cloning by limited dilution. And test by ELISA and/or FACS the supernatants to select the best producers. We do a titration with supernatants and pick the bests. > I hope this helps. > > Thanks! > > > Laura Bover, PhD > Professor > Director Monoclonal Antibody Core Facility > University of Texas M.D.Anderson Cancer Center > Associate member of The University of Texas Graduate School of Biomedical Sciences at Houston > Immunology Department/ Genomics Medicine Department > 7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039 > Houston- TX 77054 > T 713-563-2281/ 713-563-3301 > F 713-563-3276 > <email obscured> <email obscured>> > > > On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> on behalf of <email obscured>> wrote: > > Everyone in this discussion forum - Anyone have experience growing CHO cells in SS media? > > Philip, While you have them if you want to play around with culturing CHO cells in SS media that would save time. Molecular Devices and Stem Cell Technologies should have something ... maybe Fisher too. We have not, so we will need to run some pilots. > > Frances > > -----Original Message----- > From: Antibody Discussion Forum <email obscured>> on behalf of Phillip Gao <email obscured>>
> Reply-To: <email obscured>> > Date: Friday, October 12, 2018 at 2:08 PM > To: <email obscured>" <email obscured>> > Cc: Lloyd Bourne <email obscured>> > Subject: Re: [ABRF Ab Forum] ClonePix > > Hi, Frances: > > Thank you so much! > > Yes, the antibody is humanized. > > I need the PI deliver the cells and medium to me, then make a bank. I will also take a close look of the manual and related materials so that know what I am doing before I send you everything. I will defiantly test the mycoplasma before I send them to you. > > Have a nice weekend. > > Philip > > -----Original Message----- > From: Antibody Discussion Forum <email obscured>> On Behalf Of Weis-Garcia, Frances/Sloan Kettering Institute > Sent: Friday, October 12, 2018 12:55 PM > To: <email obscured> > Cc: Lloyd Bourne <email obscured>> > Subject: Re: [ABRF Ab Forum] ClonePix > > We have one, and are open to trying. > We have only grown hybridomas but if the CHO are in suspension it should be fine. > > If you send the media (liquid and semisolid) and cells, we can give it try and only charge you if it works. Fair? > > Have the cells been tested for mycoplasma? > Please confirm the antibody has a human Fc domain. > If you do not tell me the antigen, I think we can avoid signing any MTA. > > Frances > > -----Original Message----- > From: Antibody Discussion Forum <email obscured>> on behalf of Phillip Gao <email obscured>> > Reply-To: <email obscured>> > Date: Friday, October 12, 2018 at 1:10 PM > To: <email obscured>" <email obscured>> > Subject: [ABRF Ab Forum] ClonePix > > Dear Friends: > > I have a pool of mixed antibody producing CHO suspension cells. The medium is commercially available. But the problem is the yield of the antibody (IgG) is very low, <100ug/l. We have the purified antigen. > > Does anyone have a ClonePix and how much would it cost for the service? Or any other better approaches? > > Rob at Vanderbilt had one and it is no longer in service. > > Thank you for your help. > > Philip > > > Philip Gao, Ph. D. > Director, COBRE Protein Production Group > Del Shankel Structural Biology Center > 2034 Becker Drive, Room 1095 > University of Kansas > Lawrence, KS 66047 > Phone: (785) 864-1825 (office); (785) 864-3406 (lab) > Fax: (785) 864-8141 > Website: http://psf.cobre.ku.edu/cores/ppg/about > > > > ββ > View topic http://list.abrf.org/r/topic/3eLY0BxEOzs635Ju6KtFK9 > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic http://list.abrf.org/r/topic/4Cl1dkiqYfV569lP4gjVwi > Leave group <email obscured>?Subject=Unsubscribe > > > ββ > View topic http://list.abrf.org/r/topic/5iJHbs5g1ZT3bM1RD8B1SZ > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic http://list.abrf.org/r/topic/dUj44DBjpYNqPcUFvi7zG > Leave group <email obscured>?Subject=Unsubscribe > > > > The information contained in this e-mail message may be privileged, confidential, and/or protected from disclosure. This e-mail message may contain protected health information (PHI); dissemination of PHI should comply with applicable federal and state laws. If you are not the intended recipient, or an authorized representative of the intended recipient, any further review, disclosure, use, dissemination, distribution, or copying of this message or any attachment (or the information contained therein) is strictly prohibited. If you think that you have received this e-mail message in error, please notify the sender by return e-mail and delete all references to it and its contents from your systems. > > ββ > View topic http://list.abrf.org/r/topic/2nBJWp9ehr0eVk9quw6EmC > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic http://list.abrf.org/r/topic/3g3fsRz2INNio4DTv6nVle > Leave group <email obscured>?Subject=Unsubscribe >
FYI - We use anti-mouse IgG - FITC (standard secondary) from Jackson
ImmunoReagents ... and they definitely have an anti-human, so we are good.
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Laura
Bover <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 4:37 PM
To: <email obscured>" <email obscured>>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
Hi Philip,
We also have a Clonepix 2 but so far only used for hybridomas.
You will need the Detection fluorescence reagent from Molecular Devices
that will help in the selection and pick up of the human IgG good producer
clones. And I am not sure that they have a detection reagent for human IgGs. We
use the murine detection reagent only.
You may also try old fashion cloning by limited dilution. And test by ELISA
and/or FACS the supernatants to select the best producers. We do a titration
with supernatants and pick the bests.
I hope this helps.
Thanks!
Laura Bover, PhD
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Immunology Department/ Genomics Medicine Department
7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039
Houston- TX 77054
T 713-563-2281/ 713-563-3301
F 713-563-3276
<email obscured>
<email obscured>>
On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of Weis-Garcia,
Frances/Sloan Kettering Institute" <email obscured> on behalf of
<email obscured>> wrote:
Everyone in this discussion forum - Anyone have experience growing CHO
cells in SS media?
Philip, While you have them if you want to play around with culturing
CHO cells in SS media that would save time. Molecular Devices and Stem Cell
Technologies should have something ... maybe Fisher too. We have not, so we
will need to run some pilots.
Frances
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
Phillip Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 2:08 PM
To: <email obscured>" <email obscured>>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
Hi, Frances:
Thank you so much!
Yes, the antibody is humanized.
I need the PI deliver the cells and medium to me, then make a bank.
I will also take a close look of the manual and related materials so that know
what I am doing before I send you everything. I will defiantly test the
mycoplasma before I send them to you.
Have a nice weekend.
Philip
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf
Of Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, October 12, 2018 12:55 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it
should be fine.
If you send the media (liquid and semisolid) and cells, we can give
it try and only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any
MTA.
Frances
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf
of Phillip Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells.
The medium is commercially available. But the problem is the yield of the
antibody (IgG) is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the
service? Or any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
ββ
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protected health information (PHI); dissemination of PHI should comply with
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authorized representative of the intended recipient, any further review,
disclosure, use, dissemination, distribution, or copying of this message or any
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Hi Philip,
We also have a Clonepix 2 but so far only used for hybridomas.
You will need the Detection fluorescence reagent from Molecular Devices that
will help in the selection and pick up of the human IgG good producer clones.
And I am not sure that they have a detection reagent for human IgGs. We use the
murine detection reagent only.
You may also try old fashion cloning by limited dilution. And test by ELISA
and/or FACS the supernatants to select the best producers. We do a titration
with supernatants and pick the bests.
I hope this helps.
Thanks!
Laura Bover, PhD
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Immunology Department/ Genomics Medicine Department
7455 Fannin St Room 1SCR4.3015/ 1901 East Rd- Room 4SCR5.1039
Houston- TX 77054
T 713-563-2281/ 713-563-3301
F 713-563-3276
<email obscured>
<email obscured>>
On 10/12/18, 3:09 PM, "Antibody Discussion Forum on behalf of Weis-Garcia,
Frances/Sloan Kettering Institute" <email obscured> on behalf of
<email obscured>> wrote:
Everyone in this discussion forum - Anyone have experience growing CHO
cells in SS media?
Philip, While you have them if you want to play around with culturing CHO
cells in SS media that would save time. Molecular Devices and Stem Cell
Technologies should have something ... maybe Fisher too. We have not, so we
will need to run some pilots.
Frances
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
Phillip Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 2:08 PM
To: <email obscured>" <email obscured>>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
Hi, Frances:
Thank you so much!
Yes, the antibody is humanized.
I need the PI deliver the cells and medium to me, then make a bank. I
will also take a close look of the manual and related materials so that know
what I am doing before I send you everything. I will defiantly test the
mycoplasma before I send them to you.
Have a nice weekend.
Philip
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, October 12, 2018 12:55 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it
should be fine.
If you send the media (liquid and semisolid) and cells, we can give it
try and only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any
MTA.
Frances
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
Phillip Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells. The
medium is commercially available. But the problem is the yield of the antibody
(IgG) is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the
service? Or any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
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The information contained in this e-mail message may be privileged,
confidential, and/or protected from disclosure. This e-mail message may contain
protected health information (PHI); dissemination of PHI should comply with
applicable federal and state laws. If you are not the intended recipient, or an
authorized representative of the intended recipient, any further review,
disclosure, use, dissemination, distribution, or copying of this message or any
attachment (or the information contained therein) is strictly prohibited. If
you think that you have received this e-mail message in error, please notify
the sender by return e-mail and delete all references to it and its contents
from your systems.
Everyone in this discussion forum - Anyone have experience growing CHO cells in
SS media?
Philip, While you have them if you want to play around with culturing CHO cells
in SS media that would save time. Molecular Devices and Stem Cell Technologies
should have something ... maybe Fisher too. We have not, so we will need to
run some pilots.
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Phillip
Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 2:08 PM
To: <email obscured>" <email obscured>>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
Hi, Frances:
Thank you so much!
Yes, the antibody is humanized.
I need the PI deliver the cells and medium to me, then make a bank. I will
also take a close look of the manual and related materials so that know what I
am doing before I send you everything. I will defiantly test the mycoplasma
before I send them to you.
Have a nice weekend.
Philip
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, October 12, 2018 12:55 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it should be
fine.
If you send the media (liquid and semisolid) and cells, we can give it try
and only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any MTA.
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
Phillip Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells. The
medium is commercially available. But the problem is the yield of the antibody
(IgG) is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the service?
Or any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
ββ
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Hi, Frances: Let me know the cost so I can have the PI budget it assuming it will work.
Philip
-----Original Message-----
From: Gao, Philip
Sent: Friday, October 12, 2018 1:08 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: RE: [ABRF Ab Forum] ClonePix
Hi, Frances:
Thank you so much!
Yes, the antibody is humanized.
I need the PI deliver the cells and medium to me, then make a bank. I will also
take a close look of the manual and related materials so that know what I am
doing before I send you everything. I will defiantly test the mycoplasma before
I send them to you.
Have a nice weekend.
Philip
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, October 12, 2018 12:55 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it should be
fine.
If you send the media (liquid and semisolid) and cells, we can give it try and
only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any MTA.
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Phillip
Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells. The medium
is commercially available. But the problem is the yield of the antibody (IgG)
is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the service? Or
any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
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Hi, Frances: Thank you so much! Yes, the antibody is humanized. I need the PI deliver the cells and medium to me, then make a bank. I will also take a close look of the manual and related materials so that know what I am doing before I send you everything. I will defiantly test the mycoplasma before I send them to you. Have a nice weekend.
Philip
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, October 12, 2018 12:55 PM
To: <email obscured>
Cc: Lloyd Bourne <email obscured>>
Subject: Re: [ABRF Ab Forum] ClonePix
We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it should be
fine.
If you send the media (liquid and semisolid) and cells, we can give it try and
only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any MTA.
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Phillip
Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells. The medium
is commercially available. But the problem is the yield of the antibody (IgG)
is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the service? Or
any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
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We have one, and are open to trying.
We have only grown hybridomas but if the CHO are in suspension it should be
fine.
If you send the media (liquid and semisolid) and cells, we can give it try and
only charge you if it works. Fair?
Have the cells been tested for mycoplasma?
Please confirm the antibody has a human Fc domain.
If you do not tell me the antigen, I think we can avoid signing any MTA.
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Phillip
Gao <email obscured>>
Reply-To: <email obscured>>
Date: Friday, October 12, 2018 at 1:10 PM
To: <email obscured>" <email obscured>>
Subject: [ABRF Ab Forum] ClonePix
Dear Friends:
I have a pool of mixed antibody producing CHO suspension cells. The medium
is commercially available. But the problem is the yield of the antibody (IgG)
is very low, <100ug/l. We have the purified antigen.
Does anyone have a ClonePix and how much would it cost for the service? Or
any other better approaches?
Rob at Vanderbilt had one and it is no longer in service.
Thank you for your help.
Philip
Philip Gao, Ph. D.
Director, COBRE Protein Production Group
Del Shankel Structural Biology Center
2034 Becker Drive, Room 1095
University of Kansas
Lawrence, KS 66047
Phone: (785) 864-1825 (office); (785) 864-3406 (lab)
Fax: (785) 864-8141
Website: http://psf.cobre.ku.edu/cores/ppg/about
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Dear Friends: I have a pool of mixed antibody producing CHO suspension cells. The medium is commercially available. But the problem is the yield of the antibody (IgG) is very low, <100ug/l. We have the purified antigen. Does anyone have a ClonePix and how much would it cost for the service? Or any other better approaches? Rob at Vanderbilt had one and it is no longer in service. Thank you for your help. Philip Philip Gao, Ph. D. Director, COBRE Protein Production Group Del Shankel Structural Biology Center 2034 Becker Drive, Room 1095 University of Kansas Lawrence, KS 66047 Phone: (785) 864-1825 (office); (785) 864-3406 (lab) Fax: (785) 864-8141 Website: http://psf.cobre.ku.edu/cores/ppg/about
Helping each other is what this group is all about ...
So glad you are willing to drive a discussion.
See a new thread to discuss logistics.
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of "Ban,
Bhupal (bb5en)" <email obscured>>
Reply-To: <email obscured>>
Date: Monday, September 17, 2018 at 9:24 AM
To: <email obscured>" <email obscured>>
Cc: "Ban, Bhupal (bb5en)" <email obscured>>
Subject: Re: [ABRF Ab Forum] Antibody Discovery and Engineering at molecular
level
Dear Frances,
I am happy to share the ideas and challenges to discover the affinity
reagents for basic research and therapeutic application. Most importantly, how
we invest the capital and resources to get the targets as reproducible and
authenticate in this society.
I can do this discussion session on October.
Thank you for your initiation and encouraging.
With best regards,
Bhupal Ban
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, September 14, 2018 1:51 PM
To: <email obscured>
Subject: Re: [ABRF Ab Forum] Antibody Discovery and Engineering at
molecular level
Welcome Bhupal.
We are rebooting our Virtual Lab Meetings, which are casual online hours
where we bring "in" speakers, discuss papers, or can even have a brain storming
session to address long standing challenges we face in our antibody cores. It
is open to the "public" so to speak. I announce it internally here at MSKCC
and RU, as well as on this discussion forum and the ABRF Discussion Forum. May
add LinkedIn and Twitter too ... but I diverge. Would you be willing to spend
about 20 to 30 min telling us about how your core runs and the challenges you
face or advancement you are working on?
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
Bhupal <email obscured>>
Reply-To: <email obscured>>
Date: Friday, September 14, 2018 at 9:04 AM
To: <email obscured>>
Subject: [ABRF Ab Forum] Antibody Discovery and Engineering at molecular
level
Dear all group members,
Good morning
I would like to introduce myself in this group, I am core director of
Antibody Engineering and Technology core (AbET) at University of Virginia. I
am trying to explore the cutting edge technology in molecular level such and
hybridoma sequencing, humanization, affinity maturation as well as
combinatorial antibody library based antibodies discovery. Here is my core
page, link https://med.virginia.edu/antibody-engineering-technology/
Please feel free to reach out to me.
Thank you
with best regards,
Bhupal Ban, Ph.D.
Assistant Professor of Research, Cell Biology
Director of Antibody Engineering and Technology Core
University of Virginia
434-924-5379, <email obscured>
https://med.virginia.edu/antibody-engineering-technology
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Dear Frances, I am happy to share the ideas and challenges to discover the affinity reagents for basic research and therapeutic application. Most importantly, how we invest the capital and resources to get the targets as reproducible and authenticate in this society. I can do this discussion session on October. Thank you for your initiation and encouraging. With best regards, Bhupal Ban
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Friday, September 14, 2018 1:51 PM
To: <email obscured>
Subject: Re: [ABRF Ab Forum] Antibody Discovery and Engineering at molecular
level
Welcome Bhupal.
We are rebooting our Virtual Lab Meetings, which are casual online hours where
we bring "in" speakers, discuss papers, or can even have a brain storming
session to address long standing challenges we face in our antibody cores. It
is open to the "public" so to speak. I announce it internally here at MSKCC
and RU, as well as on this discussion forum and the ABRF Discussion Forum. May
add LinkedIn and Twitter too ... but I diverge. Would you be willing to spend
about 20 to 30 min telling us about how your core runs and the challenges you
face or advancement you are working on?
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Bhupal
<email obscured>>
Reply-To: <email obscured>>
Date: Friday, September 14, 2018 at 9:04 AM
To: <email obscured>>
Subject: [ABRF Ab Forum] Antibody Discovery and Engineering at molecular level
Dear all group members,
Good morning
I would like to introduce myself in this group, I am core director of
Antibody Engineering and Technology core (AbET) at University of Virginia. I
am trying to explore the cutting edge technology in molecular level such and
hybridoma sequencing, humanization, affinity maturation as well as
combinatorial antibody library based antibodies discovery. Here is my core
page, link https://med.virginia.edu/antibody-engineering-technology/
Please feel free to reach out to me.
Thank you
with best regards,
Bhupal Ban, Ph.D.
Assistant Professor of Research, Cell Biology
Director of Antibody Engineering and Technology Core
University of Virginia
434-924-5379, <email obscured>
https://med.virginia.edu/antibody-engineering-technology
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Welcome Bhupal.
We are rebooting our Virtual Lab Meetings, which are casual online hours where
we bring "in" speakers, discuss papers, or can even have a brain storming
session to address long standing challenges we face in our antibody cores. It
is open to the "public" so to speak. I announce it internally here at MSKCC
and RU, as well as on this discussion forum and the ABRF Discussion Forum. May
add LinkedIn and Twitter too ... but I diverge. Would you be willing to spend
about 20 to 30 min telling us about how your core runs and the challenges you
face or advancement you are working on?
Frances
ο»Ώ-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Bhupal
<email obscured>>
Reply-To: <email obscured>>
Date: Friday, September 14, 2018 at 9:04 AM
To: <email obscured>>
Subject: [ABRF Ab Forum] Antibody Discovery and Engineering at molecular level
Dear all group members,
Good morning
I would like to introduce myself in this group, I am core director of
Antibody Engineering and Technology core (AbET) at University of Virginia. I
am trying to explore the cutting edge technology in molecular level such and
hybridoma sequencing, humanization, affinity maturation as well as
combinatorial antibody library based antibodies discovery. Here is my core
page, link https://med.virginia.edu/antibody-engineering-technology/
Please feel free to reach out to me.
Thank you
with best regards,
Bhupal Ban, Ph.D.
Assistant Professor of Research, Cell Biology
Director of Antibody Engineering and Technology Core
University of Virginia
434-924-5379, <email obscured>
https://med.virginia.edu/antibody-engineering-technology
ββ
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Dear all group members, Good morning I would like to introduce myself in this group, I am core director of Antibody Engineering and Technology core (AbET) at University of Virginia. I am trying to explore the cutting edge technology in molecular level such and hybridoma sequencing, humanization, affinity maturation as well as combinatorial antibody library based antibodies discovery. Here is my core page, link https://med.virginia.edu/antibody-engineering-technology/ Please feel free to reach out to me. Thank you with best regards, Bhupal Ban, Ph.D. Assistant Professor of Research, Cell Biology Director of Antibody Engineering and Technology Core University of Virginia 434-924-5379, <email obscured> https://med.virginia.edu/antibody-engineering-technology
You are the best!
Frances 516-967-0656 On Thu, Sep 6, 2018, 9:29 PM David Blum <email obscured>> wrote: > Here is what we used previously. > > 1. BTX cytofusion media [gram amounts are for 500 ml of cytofusion > media] > > 1. 300 mM Sorbitol (Fisher, #BP439-500) [27.3 g] > 2. 0.1 mM Calcium Acetate (Fisher, #AC21105-2500) [.008 g or 8 mg] > 3. 0.5 mM Magnesium Acetate (Fisher, #AC42387-0050) [.0536 g or 53.6 > mg] > 4. 1.0 mg/ml BSA (Sigma, #A2153) [0.5 g] > 5. Filter sterilize and store at 4Β°C > > > On Thu, Sep 6, 2018 at 5:48 PM Weis-Garcia, Frances/Sloan Kettering > Institute <email obscured>> wrote: > > > Thanks ... I'll tickle David later unless he sees this and responds > > himself ;) > > > > You out there David ... > > > > ο»Ώ-----Original Message----- > > From: Antibody Discussion Forum <email obscured>> on behalf of > > Robert Carnahan <email obscured>> > > Reply-To: <email obscured>> > > Date: Thursday, September 6, 2018 at 5:07 PM > > To: <email obscured>" <email obscured>> > > Subject: Re: [ABRF Ab Forum] Electrofusion solution > > > > We got a recipe from David Blum and it worked great. I do not have > > the recipe here with me. > > > > Rob > > > > > > ο»ΏOn 9/6/18, 2:32 PM, "Antibody Discussion Forum on behalf of > > Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> > > on behalf of <email obscured>> wrote: > > > > Irena ... see below from the antibody discussion forum > > > > Scott ... thanks! We have another recipe we will try side by > side > > with yours should we not find a commercial source. > > > > -----Original Message----- > > From: Antibody Discussion Forum <email obscured>> on > > behalf of Scott Brown <email obscured>> > > Reply-To: <email obscured>> > > Date: Thursday, September 6, 2018 at 3:10 PM > > To: <email obscured>" <email obscured>> > > Subject: Re: [ABRF Ab Forum] Electrofusion solution > > > > β 1 file link β > > > > Hi Frances, > > I think this was the paper with the formula for the buffer > > that I used a long time ago when I first tried electrofusion. I also use > > the BTXpress medium C now, so I may try it if I run out of my supply. > > > > 280 mM inositol > > 1 mM phosphate buffer (K2HP04/KH2P04) > > 0.5 mM magnesium acetate > > 0.1 mM calcium acetate > > > > Best, > > Scott > > > > Scott A. Brown, Ph.D. > > Director of Immunologic Reagent Facility > > Department of Immunology > > 262 Danny Thomas Place > > Immunologic Reagent Facility > > Department of Immunology > > St. Jude Children's Res. Hosp. > > Memphis, TN 38105 > > w. (901) 595-4067 > > F. (901) 595-3107 > > c. (901) 674-9960 > > > > > > > > > > > > > > -----Original Message----- > > From: Antibody Discussion Forum <email obscured>> On > > Behalf Of Frances Weis-Garcia > > Sent: Thursday, September 06, 2018 1:08 PM > > To: <email obscured> > > Subject: [ABRF Ab Forum] Electrofusion solution > > > > For those who do electrofusion, we are looking for another > > source of electrofusion solution/media - the solution in which the cells > > are suspended for the fusion. > > > > We have used BTXpress Cytofusion Medium C but they are not > > back ordered until November ... ugh! > > > > > > > https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.btxonline.com%2Faccessories%2Fbuffers%2Fbtxpress-cytofusion-medium-c-500-ml-volume.html&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=7b7eKH%2BKs9klucdU1WouxVju9MAY5gY8kZkh8pPXyUw%3D&reserved=0 > > > > Thanks in advance for the help. > > > > ββ > > View topic > > > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F4ihjgmvh7vhGv0Y45ln2Xt&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=1xx1%2F%2B6jGUsDNnXXN%2F%2Bwk0aS27qmInmy7lfaMlAMzw0%3D&reserved=0 > > Leave group <email obscured> > ?Subject=Unsubscribe > > > > > > ________________________________ > > > > Email Disclaimer: > > > https://na01.safelinks.protection.outlook.com/?url=www.stjude.org%2Femaildisclaimer&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=dSTvtQ6SKzQj37EQNW4w%2FgCAHmzhwu1xfvhHjoKFXR4%3D&reserved=0 > > Consultation Disclaimer: > > > https://na01.safelinks.protection.outlook.com/?url=www.stjude.org%2Fconsultationdisclaimer&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=SSnKI23ZXq9I19tqGLu%2FKYa%2FbTxiKjbY6gbVnbXpfHQ%3D&reserved=0 > > > > β 1 file β > > > > β‘ Vienken_et_al-1985-FEBS_Letters.pdf (303kb) > > > > > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ffile%2F8ngyl27RFNkjHBbyhUbIXG7ZzQw-1gOb-2HGLtwq%2F&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=%2FR9dh5BDbCX31xOF8g%2BG2VjOo3Wbm%2Bz6c8BGqGwPhOU%3D&reserved=0 > > > > ββ > > View topic > > > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F6QZNcpjy0EuFY0QIk1JQdA&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=jC%2FHrTAJlKkAzvpslVbyx0acwBgcTwFmAriOFQx3q9U%3D&reserved=0 > > Leave group <email obscured> > ?Subject=Unsubscribe > > > > > > > > > > > > ββ > > View topic > > > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F7InAyKlKu0o92V4ujSV6Tj&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=RaWu2RJ9d9SCyrHe9b3QLSLOI%2FwNY%2B9ujTsW89Zhpb0%3D&reserved=0 > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > ββ > > View topic http://list.abrf.org/r/topic/G2xK7o9JSvGbxsmSqj38R > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > > > ββ > > View topic http://list.abrf.org/r/topic/K7A9heUhQsWS0aGa7J1Qq > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic http://list.abrf.org/r/topic/1cl23fbPI4H5JTgg6VCHV8 > Leave group <email obscured>?Subject=Unsubscribe > >
Here is what we used previously.
1. BTX cytofusion media [gram amounts are for 500 ml of cytofusion
media]
1. 300 mM Sorbitol (Fisher, #BP439-500) [27.3 g]
2. 0.1 mM Calcium Acetate (Fisher, #AC21105-2500) [.008 g or 8 mg]
3. 0.5 mM Magnesium Acetate (Fisher, #AC42387-0050) [.0536 g or 53.6
mg]
4. 1.0 mg/ml BSA (Sigma, #A2153) [0.5 g]
5. Filter sterilize and store at 4Β°C
On Thu, Sep 6, 2018 at 5:48 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks ... I'll tickle David later unless he sees this and responds > himself ;) > > You out there David ... > > ο»Ώ-----Original Message----- > From: Antibody Discussion Forum <email obscured>> on behalf of > Robert Carnahan <email obscured>> > Reply-To: <email obscured>> > Date: Thursday, September 6, 2018 at 5:07 PM > To: <email obscured>" <email obscured>> > Subject: Re: [ABRF Ab Forum] Electrofusion solution > > We got a recipe from David Blum and it worked great. I do not have > the recipe here with me. > > Rob > > > ο»ΏOn 9/6/18, 2:32 PM, "Antibody Discussion Forum on behalf of > Weis-Garcia, Frances/Sloan Kettering Institute" <email obscured> > on behalf of <email obscured>> wrote: > > Irena ... see below from the antibody discussion forum > > Scott ... thanks! We have another recipe we will try side by side > with yours should we not find a commercial source. > > -----Original Message----- > From: Antibody Discussion Forum <email obscured>> on > behalf of Scott Brown <email obscured>> > Reply-To: <email obscured>> > Date: Thursday, September 6, 2018 at 3:10 PM > To: <email obscured>" <email obscured>> > Subject: Re: [ABRF Ab Forum] Electrofusion solution > > β 1 file link β > > Hi Frances, > I think this was the paper with the formula for the buffer > that I used a long time ago when I first tried electrofusion. I also use > the BTXpress medium C now, so I may try it if I run out of my supply. > > 280 mM inositol > 1 mM phosphate buffer (K2HP04/KH2P04) > 0.5 mM magnesium acetate > 0.1 mM calcium acetate > > Best, > Scott > > Scott A. Brown, Ph.D. > Director of Immunologic Reagent Facility > Department of Immunology > 262 Danny Thomas Place > Immunologic Reagent Facility > Department of Immunology > St. Jude Children's Res. Hosp. > Memphis, TN 38105 > w. (901) 595-4067 > F. (901) 595-3107 > c. (901) 674-9960 > > > > > > > -----Original Message----- > From: Antibody Discussion Forum <email obscured>> On > Behalf Of Frances Weis-Garcia > Sent: Thursday, September 06, 2018 1:08 PM > To: <email obscured> > Subject: [ABRF Ab Forum] Electrofusion solution > > For those who do electrofusion, we are looking for another > source of electrofusion solution/media - the solution in which the cells > are suspended for the fusion. > > We have used BTXpress Cytofusion Medium C but they are not > back ordered until November ... ugh! > > > https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.btxonline.com%2Faccessories%2Fbuffers%2Fbtxpress-cytofusion-medium-c-500-ml-volume.html&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=7b7eKH%2BKs9klucdU1WouxVju9MAY5gY8kZkh8pPXyUw%3D&reserved=0 > > Thanks in advance for the help. > > ββ > View topic > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F4ihjgmvh7vhGv0Y45ln2Xt&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=1xx1%2F%2B6jGUsDNnXXN%2F%2Bwk0aS27qmInmy7lfaMlAMzw0%3D&reserved=0 > Leave group <email obscured>?Subject=Unsubscribe > > > ________________________________ > > Email Disclaimer: > https://na01.safelinks.protection.outlook.com/?url=www.stjude.org%2Femaildisclaimer&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=dSTvtQ6SKzQj37EQNW4w%2FgCAHmzhwu1xfvhHjoKFXR4%3D&reserved=0 > Consultation Disclaimer: > https://na01.safelinks.protection.outlook.com/?url=www.stjude.org%2Fconsultationdisclaimer&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=SSnKI23ZXq9I19tqGLu%2FKYa%2FbTxiKjbY6gbVnbXpfHQ%3D&reserved=0 > > β 1 file β > > β‘ Vienken_et_al-1985-FEBS_Letters.pdf (303kb) > > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ffile%2F8ngyl27RFNkjHBbyhUbIXG7ZzQw-1gOb-2HGLtwq%2F&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=%2FR9dh5BDbCX31xOF8g%2BG2VjOo3Wbm%2Bz6c8BGqGwPhOU%3D&reserved=0 > > ββ > View topic > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F6QZNcpjy0EuFY0QIk1JQdA&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=jC%2FHrTAJlKkAzvpslVbyx0acwBgcTwFmAriOFQx3q9U%3D&reserved=0 > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F7InAyKlKu0o92V4ujSV6Tj&data=02%7C01%7Crobert.carnahan%40vanderbilt.edu%7C5a0c356354924f80034b08d6142f7db2%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636718591530599883&sdata=RaWu2RJ9d9SCyrHe9b3QLSLOI%2FwNY%2B9ujTsW89Zhpb0%3D&reserved=0 > Leave group <email obscured>?Subject=Unsubscribe > > > > > ββ > View topic http://list.abrf.org/r/topic/G2xK7o9JSvGbxsmSqj38R > Leave group <email obscured>?Subject=Unsubscribe > > > > > > ββ > View topic http://list.abrf.org/r/topic/K7A9heUhQsWS0aGa7J1Qq > Leave group <email obscured>?Subject=Unsubscribe > >