Hi,
There are 2 options during the installation process: Regulatory Mode or
non-Regulatory Mode.
If you chose non-regulatory mode, you do not have to continually log into the
program BUT you are unable to do all of the software functions / changes. This
may be your problem.
If you chose the Regulatory mode, you should have access to all functions but
you'll have to relog-in every 99 minutes or less.
Unfortunately, to run your sequencer you pretty much need the Regulatory Mode.
I hope this solves your issues.
David Chiara
Neurobiology
Harvard Medical School
Rest of post
-----Original Message-----
From: ABRF Discussion Forum <email obscured>> On Behalf Of Pawel Mak
Sent: Friday, September 21, 2018 4:59 AM
To: <email obscured>
Subject: [ABRF Discussion Forum] Procise protein sequencer PC software problem
Dear Procise users
Due to problems with my old-fashioned Power Macintosh computer I have recently
bought Procise PC v2.1 software and installed it on a Win 2000 computer.
Everything works fine with two exceptions - the panel "Pressures &
Temperatures" does not work (the communication "Operation not supported"
appears), I am also unable to open the "Preferences" window.
I would be grateful for your help.
Regards,
Pawel Mak
--
Dr hab. Paweł Mak
Zakład Biochemii Analitycznej
Wydział Biochemii, Biofizyki i Biotechnologii
Uniwersytet Jagielloński
Ul. Gronostajowa 7, 30-387 Kraków
Pawel Mak, PhD, DSc
Department of Analytical Biochemistry
Faculty of Biochemistry, Biophysics and Biotechnology
Jagiellonian University
Gronostajowa 7 St., 30-387 Krakow, POLAND
e-mail: <email obscured>
www: http://www.wbbib.uj.edu.pl
www: http://www.zba.wbbib.uj.edu.pl/
www: http://www.mcb.uj.edu.pl
Fax +48 12 6646915
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Hello all,
I’m looking into using a fluorometric peptide assay to measure samples with low
concentration of peptides after trypsin digest and cleanup. I have been using
A205 (Scopes) but it is too variable because at 205nm there are too many
interfering substances that will absorb at that end of the spectrum.
The Pierce Fluorometric assay seems ideal since it reacts with the peptide NH2
and is somewhat more sensitive than the Nano Orange reagent from Molecular
Devices. I would really appreciate getting some feedback from those who use
fluorometric assays for measuring peptide concentrations or other assays that
are sensitive enough that you don’t end up consuming a large portion of your
precious sample just to measure its concentration.
Thanks in advance,
Brian
Brian Hampton
Program Director
Center for Vascular and Inflammatory Diseases
University of Maryland School of Medicine
BRB Room 7-018
655 West Baltimore St
Baltimore MD. 21201
Dear Procise users
Due to problems with my old-fashioned Power Macintosh computer I have
recently bought Procise PC v2.1 software and installed it on a Win 2000
computer. Everything works fine with two exceptions - the panel
"Pressures & Temperatures" does not work (the communication "Operation
not supported" appears), I am also unable to open the "Preferences" window.
I would be grateful for your help.
Regards,
Pawel Mak
Rest of post
--
Dr hab. Paweł Mak
Zakład Biochemii Analitycznej
Wydział Biochemii, Biofizyki i Biotechnologii
Uniwersytet Jagielloński
Ul. Gronostajowa 7, 30-387 Kraków
Pawel Mak, PhD, DSc
Department of Analytical Biochemistry
Faculty of Biochemistry, Biophysics and Biotechnology
Jagiellonian University
Gronostajowa 7 St., 30-387 Krakow, POLAND
e-mail: <email obscured>
www: http://www.wbbib.uj.edu.pl
www: http://www.zba.wbbib.uj.edu.pl/
www: http://www.mcb.uj.edu.pl
Fax +48 12 6646915
Thank you KTN. Appreciate your time.
The MPC ClientPro 565 computer on our Shimadzu Prominence HPLC also just went
dead. Does anyone have one of these lying around, or know where we can either
fix ours or pick up a second hand one?
Any and all advice will be appreciated
Kym
Kym Francis Faull, Ph.D.
Director, Pasarow Mass Spectrometry Laboratory
Semel Institute of Neuroscience and Human Behavior
Professor, Department of Psychiatry and Biobehavioral Sciences
David Geffen School of Medicine at UCLA
Telephone: (310) 206 7881 (office)
Telephone: (310) 206 7886 (laboratory)
Facsimile: (310) 206 2161
Electronic mail: <email obscured> <email obscured>>
Believe it or not but we still use an HP 1090 Chemstation as a stand-alone HPLC
. However, we need a back-up computer for this unit as concerns are mounting
that the existing computer is going to fail soon. Does anyone have the
computer that was attached to one of these systems? We are prepared to pay for
shipping if you would like to donate the unit.
Kym
Kym Francis Faull, Ph.D.
Director, Pasarow Mass Spectrometry Laboratory
Semel Institute of Neuroscience and Human Behavior
Professor, Department of Psychiatry and Biobehavioral Sciences
David Geffen School of Medicine at UCLA
Telephone: (310) 206 7881 (office)
Telephone: (310) 206 7886 (laboratory)
Facsimile: (310) 206 2161
Electronic mail: <email obscured> <email obscured>>
Hi ABRF Protein Sequencers,
We developed 2 contaminating peaks on our chromatograms occurring just before
Phe (Large) and after Leu (small) that were due to a bad lot of R4A.
The Lot # is 18D010. ThermoFisher is aware of the issue. Thought you should
be too.
Sincerely,
David Chiara
Neurobiology
Harvard Medical School
My group is two, myself (director) and a technician. I expect my tech to
order consumables, maintain the inventory of the lab, handle the prep for
proteomics samples (with my input first), handle small molecule analysis
(with my input if things aren't working), return small molecule data to
investigators, and do routine lab maintenance. Billing, instrument
repairs, proteomics sample analysis, data analysis, project discussion with
investigators ... these are all my own duties.
Cheers,
KTN
On Thu, Sep 20, 2018 at 5:36 AM, Hampton, Brian <email obscured>>
wrote:
Rest of post
> Just an FYI, new subscribers to the ABRF forums are moderated initially to
> reduce spammers from joining. Until moderation is removed from your
> account, your posts will be withheld from the list. However, the system is
> not perfect, so if you have already posted to e.g the CANCC list before I
> removed moderation (which I just did), you may have to send that post again.
>
>
> Cheers,
>
>
> Brian
>
> ________________________________
> From: ABRF Discussion Forum <email obscured>> on behalf of kgij2 <
> <email obscured>>
> Sent: Thursday, September 20, 2018 3:37:52 AM
> To: <email obscured>
> Subject: Re: [ABRF Discussion Forum] Duties of A Chief Research Lab
> Tecnician in Proteomics
>
> CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS email
> system. Whether the sender is known or not known, hover over any links
> before clicking and use caution opening attachments.
>
>
>
> Thanks Brian. I really need few more comments. Good advice.
>
> Best wishes!
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>
> ――
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>
--
Kristina T. Nelson, Ph.D.
Director
Chemical and Proteomic Mass Spectrometry Core Facility
Oliver Hall Education Wing, Room 1077
1001 W. Main Street
Richmond, VA 23284
804-828-7445
Just an FYI, new subscribers to the ABRF forums are moderated initially to
reduce spammers from joining. Until moderation is removed from your account,
your posts will be withheld from the list. However, the system is not perfect,
so if you have already posted to e.g the CANCC list before I removed moderation
(which I just did), you may have to send that post again.
Rest of post
Cheers,
Brian
________________________________
From: ABRF Discussion Forum <email obscured>> on behalf of kgij2
<email obscured>>
Sent: Thursday, September 20, 2018 3:37:52 AM
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] Duties of A Chief Research Lab Tecnician
in Proteomics
CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS email
system. Whether the sender is known or not known, hover over any links before
clicking and use caution opening attachments.
Thanks Brian. I really need few more comments. Good advice.
Best wishes!
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Hi Adam,
We also had this issue time and again with the EasySpray and eventually stopped
using them. It was quite recently. We made a huge effort to troubleshoot and
Thermo helped us a lot but at the end of the day they were just not performing.
Best,
Yishai
> On 12 Sep 2018, at 19:44, Adam Rabalski <email obscured>> wrote:
>
> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
Rest of post
>
> Best,
>
> Adam
> ――
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> Leave group <email obscured>?Subject=Unsubscribe
>
Thanks Brian. I really need few more comments. Good advice.
Best wishes!
Hi,
You might get additional responses by posting to the ABRF Core Administrator's
forum. That list discusses wide ranging administrative topics concerning core
facilities. See: http://list.abrf.org/groups/cancc/
Rest of post
Cheers,
Brian
________________________________
From: ABRF Discussion Forum <email obscured>> on behalf of kgij2
<email obscured>>
Sent: Wednesday, September 19, 2018 4:33:52 AM
To: <email obscured>
Subject: [ABRF Discussion Forum] Duties of A Chief Research Lab Tecnician in
Proteomics
CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS email
system. Whether the sender is known or not known, hover over any links before
clicking and use caution opening attachments.
Hi all,
I would really appreciate as many replies as possible to receive on this from
those who are in charge of proteomics labs..
What should be the duties of a chief research lab technician in a proteomics
lab be in a postgraduate institution? It is a group of three. A
bioinfamitician, the proteomics group manager and the Technician recruited as
the Chief Research Lab Tecnician in Proteomics.
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Hi,
keeping the instruments running, taking care of spare parts and consumables,
columns, solvents, standards, running the standards and doing calibration and
tuning of the instruments, maintaining HPLC/UHPLC systems, keeping book about
this, communicating with service engineers and suppliers, running the samples,
evaluating the instrument performance, decides to run samples which are not ok
(with talking to the manager). Looking after the computers and the storage,
shares with the bioinformatician even it is not the job of any of these, but it
is a small group so everyone has to take several hats. Also communication with
customers together with the rest of the group. Telling people how to do sample
prep, what are common mistakes and how to do it better. In the end the week has
only 7 days and the day only 24 h, it is a full time job, if things run fine
and nothing urgent to do, read literature and learn about new developments. I
always allowed the technicians to go to company courses, not because of free
food (, and if possible also to conferences. Important always keep them in any
discussion, they are not only taking orders, they are the people doing the
work. But even as group manager I never could keep my fingers away from real
work. And inform them about the findings the bioinformatics person made, always
in the loop.
Just my 5 cents ---
Manfred
Am 19.09.18, 10:33 schrieb "ABRF Discussion Forum im Auftrag von kgij2"
<email obscured> im Auftrag von <email obscured>>:
Hi all,
I would really appreciate as many replies as possible to receive on this
from those who are in charge of proteomics labs..
What should be the duties of a chief research lab technician in a
proteomics lab be in a postgraduate institution? It is a group of three. A
bioinfamitician, the proteomics group manager and the Technician recruited as
the Chief Research Lab Tecnician in Proteomics.
――
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Leave group <email obscured>?Subject=Unsubscribe
Hi all,
I would really appreciate as many replies as possible to receive on this from
those who are in charge of proteomics labs..
What should be the duties of a chief research lab technician in a proteomics
lab be in a postgraduate institution? It is a group of three. A
bioinfamitician, the proteomics group manager and the Technician recruited as
the Chief Research Lab Tecnician in Proteomics.
Dear all,
I am looking Flp-In™-293 cell line and pOG44 plasmid DNA as cotransfection. if
you have, please let me know.
I appreciate your help.
thank you
Dear all,
as I can see there is some interest in µPAC. I have a presentation on
that and can load it on some place for download. Please give me a day or
two to figure out how to do that since I am at
https://reduse2018.meduniwien.ac.at/ and have no direct access to my PC.
After that, we can discuss all you are interested about. There are many
results I have generated with this column, 2 m flow rates of 800 nl/min
and 600 nl/min, respectively. All samples analyzed, so far are of human
origin - serum, urine, tissue. The column enables better sequence
coverage as compared to the PepMap 50 cm and, simultaneously, higher ID
rates. As I wrote, I have a lot of material but I cannot discuss all of
it due to fact that I do not "own" the data and I need to ask for
permission on showing the most complex ones. You might want to contact
me privately for detailed description of the methods used.
Many greetings
Goran
On 13-Sep-18 4:40 PM, Adam Rabalski wrote:
> Hi Duncan,
>
> Yes that's correct I'm using a vented trap configuration. The spray current
is about 0.05-0.10 uA when I observe sputtering. The voltage is off during the
trapping phase as the instrument is in standby. I'll add in an additional
segment at the beginning of the method to turn the spray voltage off for a few
minutes after valve switching as you suggest.
>
> Thanks,
>
> Adam
>
> PS
>
> Goran, I'm interested in seeing your setup as well! What length of the uPAC
do you use and what are your ID rates like for a complex sample such as
mammalian proteome?
Rest of post
> ――
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>
--
Goran Mitulović, Ph.D.
ORCID 0000-0003-1964-3965
Head Proteomics Core Facility
Medical University of Vienna
Clinical Department of Laboratory Medicine
Proteomics Core Facility, E5, LS5H
Waehringer Guertel 18-20
A-1090 Wien, Austria
Tel: +43 1 40400 53750
Mail: <email obscured>
Web: www.meduniwien.ac.at/proteomics-core
Hi Duncan,
Yes that's correct I'm using a vented trap configuration. The spray current is
about 0.05-0.10 uA when I observe sputtering. The voltage is off during the
trapping phase as the instrument is in standby. I'll add in an additional
segment at the beginning of the method to turn the spray voltage off for a few
minutes after valve switching as you suggest.
Thanks,
Adam
PS
Goran, I'm interested in seeing your setup as well! What length of the uPAC do
you use and what are your ID rates like for a complex sample such as mammalian
proteome?
Hello Adam
I was referring to the ion spray current read back not the TIC. The current
read back will allow you to see if there is an issue that correlates with the
spitting.
If you are using a nAcquity with a trap I'm guessing you are using a vented
configuration. Is this the case? If so, this involves having no flow through
the analytical column, liquid junction and emitter whilst you load the sample.
If you are applying HV during this phase of the method, thats not good for the
EasySpray liquid junction or emitter. When I used a vented n Acquity and easy
spray setup I turned off the HV during the loading phase and only introduced it
a couple of minutes after the valve switched and flow was returned to the
analytical column. this means you only have HV supply whilst the liquid
junction is fully flowing. I found this massively improved EasySpray life span
and eliminated spitting all together.
Hope this helps
Rest of post
Duncan
________________________________________
From: ABRF Discussion Forum <email obscured>] on behalf of Adam Rabalski
<email obscured>]
Sent: 12 September 2018 19:52
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] EASY-Spray C18 PepMap RSLC Emitter Spray
Sputter
Hi Duncan,
I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA). I’ll
have to check what signal I have on a non-sputtering column. I’m running at 2.2
kV spray voltage. It’s a Waters NanoAcquity UPLC.
Thanks,
Adam
> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>>
wrote:
>
> Hello Adam
>
> Do you see any ion spray current difference when spluttering versus spraying?
What ion spray voltage are you using? Are you using an EasynLC or something
else?
>
>
>
> Cheers
>
> Duncan
>
> Sent from my iPhone
>
>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>
>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
>>
>> Best,
>>
>> Adam
>> ――
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>>
>
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Thank you very much Goran, I'd love to see pictures of your setup.
Bill
On Thu, Sep 13, 2018 at 11:27 AM, Goran Mitulović <
Rest of post
<email obscured>> wrote:
> Actually, you can use them as a normal nano column. There are no
> integrated emitters and you have to use either steel or glass emitters. I
> am very happy with stainless steel emitters (ES542) from Thermo and their
> classic nano-spray source, and, if you wish, I can send you some pictures
> of the setup and chromatograms.
>
> Goran
>
>
>
>
>
> On 13-Sep-18 10:15 AM, William Breuer wrote:
>
>> that's a very interesting suggestion - are the PharmaFluidics columns
>> built
>> to fit into Thermo's Nano-spray source?
>>
>> On Thu, Sep 13, 2018 at 11:08 AM, Goran Mitulović <
>> <email obscured>> wrote:
>>
>> Try the µPAC columns from PharmaFluidics. These are great with very low
>>> back pressure and low background and the separation of peptides is
>>> superior
>>> as compared to any packed column.
>>>
>>> Success
>>>
>>> Goran
>>>
>>>
>>>
>>> On 13-Sep-18 9:21 AM, William Breuer wrote:
>>>
>>> We have also noticed a significant deterioration in the quality of spray
>>>> and lifetime of the PepMap columns from Thermo. The proportion of duds
>>>> that
>>>> die within a couple of weeks has been increasing alarmingly.
>>>> Bill Breuer
>>>>
>>>> On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>>
>>>> wrote:
>>>>
>>>> Hi Duncan,
>>>>
>>>>> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1%
>>>>> FA).
>>>>> I’ll have to check what signal I have on a non-sputtering column. I’m
>>>>> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>>>>>
>>>>> Thanks,
>>>>>
>>>>> Adam
>>>>>
>>>>> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>.
>>>>> uk
>>>>> wrote:
>>>>>
>>>>> Hello Adam
>>>>>>
>>>>>> Do you see any ion spray current difference when spluttering versus
>>>>>>
>>>>>> spraying? What ion spray voltage are you using? Are you using an
>>>>> EasynLC
>>>>> or
>>>>> something else?
>>>>>
>>>>>
>>>>>> Cheers
>>>>>>
>>>>>> Duncan
>>>>>>
>>>>>> Sent from my iPhone
>>>>>>
>>>>>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>>>>>
>>>>>>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
>>>>>>>
>>>>>>> (ES802) column. We have an inline two-column system with trapping
>>>>>>
>>>>> column.
>>>>> We've been diligent in using detergent clean up kits when needed and
>>>>> perform SPE on all samples. The set of samples where spray sputtering
>>>>> appeared were SCX fractions from a StageTip SCX fractionation approach
>>>>> (using ammonium acetate buffer). The column had 15 injections prior to
>>>>> this
>>>>> of which half were an E. coli peptide digest and the other half were
>>>>> acetonitrile ("blank") injections. Has anyone experienced spray
>>>>> sputtering
>>>>> issues with the Easy-Spray columns? In general what do you when you
>>>>> encounter spray sputtering ("solvent beading") at the emitter? Any
>>>>> suggestions for cleaning solutions other than high organic to flush the
>>>>> column?
>>>>>
>>>>> Best,
>>>>>>
>>>>>>> Adam
>>>>>>> ――
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>>>>>>>
>>>>>>> ――
>>>>>>>
>>>>>> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
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>>>>>>
>>>>>> ――
>>>>>>
>>>>> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
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>>>>>
>>>>>
>>>>> ――
>>>>>
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>>>>
>>>>
>>>> --
>>>
>>>
>>> Goran Mitulović, Ph.D.
>>> ORCID 0000-0003-1964-3965
>>> Head Proteomics Core Facility
>>> Medical University of Vienna
>>> Clinical Department of Laboratory Medicine
>>> Proteomics Core Facility, E5, LS5H
>>> Waehringer Guertel 18-20
>>> A-1090 Wien, Austria
>>> Tel: +43 1 40400 53750
>>> Mail: <email obscured>
>>> Web: www.meduniwien.ac.at/proteomics-core
>>>
>>>
>>> ――
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>>>
>>>
>>> ――
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>>
>>
> --
>
>
> Goran Mitulović, Ph.D.
> ORCID 0000-0003-1964-3965
> Head Proteomics Core Facility
> Medical University of Vienna
> Clinical Department of Laboratory Medicine
> Proteomics Core Facility, E5, LS5H
> Waehringer Guertel 18-20
> A-1090 Wien, Austria
> Tel: +43 1 40400 53750
> Mail: <email obscured>
> Web: www.meduniwien.ac.at/proteomics-core
>
>
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>