I spin down my cells with PBS, then discard the PBS and add RIPA lysis buffer to lyse them. However, I always have foam/ a lot of bubbles forming whenever I add the lysis buffer and resuspend my cells to mix them. Will this affect my western blot results? Will this cause my western blots to have zero results i.e totally no protein band formation?
Shin Murakami
Lysis buffer has detergent in it and, if you mix it, it will make bubbles. It is similar when you wash your hands with soap. I would suggest mixing the buffer gently using the pipetman but you need to make sure the cells are lysed well.
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