How much concentration of CHAPS can i use in lysis buffer？
I would like to purify protein. But only CHAPS is compatible with mass spectrometry.
http://agscientific.com/blog/index.php/2012/10/01/chaps14-faqs-protocols-you-must-have/
may be useful
Well compatible is a big word.... You will be stuck with a big peak around 615amu at one point in your LC_MS trace. The amount needed depend on the type of cell you are using, try different concentration and verify if your cells are lysed. I would be inclined to precipitate after (using either acetone or TCA or chlorof/Metoh) to remove your detergent. You can also remove chaps if you clean your tryptic samples on a SCX column (make sure you have a low concentration of ABC during digest).
good luck!
T
@ Liming Zhang , I used CHAPS in my proteomics research. Before LC-MS/MS (Agilent Technologies), I practise to de-salt the protein samples using C18 column. The outcomes were really good.
For SH-SY5Y cells, I used 2% CHAPS. In my proteomic lab, we find this concentration optimal. You may give a try. All best luck!
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