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Low pH Cell Lysis Buffer

cell lysis ph SDS-PAGE

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#1 ahitron

ahitron

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Posted 02 January 2013 - 01:13 PM

Hi all, I'm using a dimedone-based probe to tag proteins with specific cysteine modifications. An article I recently came across that looked at the binding efficiency of this tag under different pH, and they found that the probe was much more efficient at binding to sulfenic acid-cysteines at low pH (~5.5). Since the cysteine sulfenic acid is already very labile, I want to make sure that I have the most efficient binding conditions I can, but I'm a little concerned about using a mildly acidic lysis buffer. So, I was wondering if anyone had any advice on the use of low-pH cell lysis conditions. Would the low pH hurt my lysis, or alter the composition of the lysate (aside from possible denaturation)?





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