Post in SDC precipitation issue

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From: ABRF Discussion Forum <email obscured>> on behalf of Chen, Zhuo, Dr.
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Sent: 09 March 2022 12:07
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Subject: [EXTERNAL] Re: [ABRF Discussion Forum] SDC precipitation issue

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Dear Eric,
Great tip! Indeed, my TECP solution was apparently acidic and after adjusting
the pH, it is all clear now. Thank you so much!

Zhuo

-----Original Message-----
From: ABRF Discussion Forum <email obscured>> On Behalf Of Anderson, David
Eric (NIH/NIDDK) [E]
Sent: 09 March 2022 17:59
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] SDC precipitation issue

My guess is you are too acidic - check the pH of the TCEP solution and if it is
acidic that would be consistent. If that is the case  you can try slowly moving
the pH up by adding tiny amounts of base until it clears. It is also possible
it is specific to the TCEP addition (I have little experience with TCEP and
soaps but some things do specific aggregation).

Eric

D. Eric Anderson,    Ph.D
Advanced Mass Spectrometry Core Facility, NIDDK NIH


Building 8 Room B2A19, 9000 Rockville Pike, Bethesda MD, 20892 USA
(301)-496-7546



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From: ABRF Discussion Forum <email obscured>> on behalf of Chen, Zhuo, Dr.
<email obscured>>
Sent: 09 March 2022 11:53
To: <email obscured>
Subject: [EXTERNAL] [ABRF Discussion Forum] SDC precipitation issue

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Hello all,
I just start to try digestion with SDC, I resuspended my protein pellet
(Acetone precipitated) with 1% SDC in 50 mM HEPES buffer (pH 7.5) and when I
add TCEP to 10 mM, my solution turned total milky. I have not clue what is
going on. Should I use a different buffer instead of HEPES?

Thank you!

Zhuo Chen

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