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WHOLE CELL PROTEIN EXTRACTS
This if for both cytoplasmic and nuclear proteins-if you want only nuclear follow nuclear extract protocol.
20 mM Tris pH 7.5
100 mM NaCl
0.5% NP-40
0.5 mM EDTA
0.5 mM PMSF
0.5% protease inhibitor cocktail (Sigma)
pipet cells up and down extensively and then sit on ice for 10 minutes.
ALTERNATIVELY-FOR VERY INSOLUBLE (i.e. cytoskeletal proteins etc.)---
Cell extraction buffer
20 mM Tris pH8.0
100 mM NaCl
1 mM EDTA
0.1% SDS
1 mM PMSF
1X protease inhibitor cocktail