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for protein interaction what lysis buffer I should use? - (Apr/11/2006 )

Dear all,
I will so some protein interaction work finally. actrully I want to know what things can not use and will disturb the protein interaction? so I will know what I can use for lysis and so on to finally protein interaction.

Thanks a milion


it depends on the type of protein interactions. I've had greater success with a .5% Triton buffer than with something stronger like RIPA.
"F" buffer

10mM Tris ph 7.05 form 1M stock
50mM NaCl from 5M stock
39mM Na4pyrophosphate (3.99 gm)
50mM NaF 1M stock
5uM ZnCl2 from 1M stock
10% glycerol
0.5% Triton X 100
pH to 7.05
I add a protease inhibitor cocktail