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NP 40 lysis buffer


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#1 bugsbunny66632

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Posted 09 July 2011 - 10:58 AM

Hi,

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?

Thanks!

#2 bob1

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Posted 10 July 2011 - 04:37 PM

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.




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