3T3 cells and lysis buffer recepie - western blot (Aug/21/2006 )

Hi
I am working on 3T3 cells, could any body tell me exact lysis buffer receipe of protein extraction for western blots, as by veiwng litrature i found variety.
Looking forward for prompt assisitance.
Thanks

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This depends on what you would like to do with your protein after you lyse the cells. A good one is:
-150mM NaCl
-10% glycerol
-1% NP-40
-5mM MgCl2 (if you need it)
-protease inhibitors.

This is good for WB and also for coIP, but forr coIP you should find the right salt concentration

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