NP 40 lysis buffer - (Jul/09/2011 )
I am making NP40 lysis buffer. To make 10% or 1% NP40 - do I just dilute the detergent in water? Also - any thoughts on how to make sodium vanadate?
Usually the detergent is diluted in a buffer of some sort containing salts and something to maintain the pH as part of the lysis buffer. You could dilute it in water first and then add to a sightly more concentrated buffer so as to bring to the correct concentrations if you wished, but it seems slightly counterproductive.
For orthovanadate preparation, see this paper: Gordon, J.A. (1991) Methods in Enzymology, 201, 477-482.