? on protein extraction

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Tue Jun 22 12:19:03 EST 1999


In article <376FA782.6A0E75E0 at pitt.edu>, Ruben Zamora <zamorar at pitt.edu> wrote:
:Hello,
:
:I would like to isolate protein from cultured cells to perform Western
:blot. What is the best method for extraction, to lyse the cells by
:adding Triton X-100 to the buffer or just leaving Triton X-100 out the
:extraction buffer and completing 4 or 5 freeze-thaw cycles?
:
If you don't care about your protein's activity, then it does not matter.
The only advantage of freeze/thaw is that you are not going to get 
membrane proteins into solution. 

I like Triton lysis because it is very fast and reproducible between 
samples.

        - Dima

        



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