wash buffer vs lysis buffer PH 7.4 or 8.0 - (Dec/05/2006 )
In some protocols for IP, people just use lysis buffer to wash, so the wash buffer is just the lysis buffer. In some protocals, the wash buffer is TBS, doen not contain detergent as lysis buffer,
also the PH of lysis buffer (tris-cl) is also different from protocals to protocals,some 7.4, 7.5 , some 8.0? Is there any difference?
usually protease inhibitor is added to the lysis buffer, should it also be added to the wash buffer(if it's not the same as lysis buffer) and elution buffer?
I use pH 7.4 for both lysis (Tris-HCl) and wash (HEPES) buffers. I don't know about other pH... I mean, pH 7.4 is physiological pH, isn't it? For wash buffer I like using more ionic strength, so it's not the same buffer. In wash buffer I do not use protease inhibitors and haven't any problem (only a phosphatase inhibitor according my goal).