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Lysis buffer with 1% tritonX - Kindly sugget me its other ingredients (Jan/19/2006 )

Hello friends, iam trying to isolate proteins from H9 cell line using lysis bufer containing 1% triton x .
i would like to know what other ingrecients should this lysis buffer contain. and i would also like to know what role does each of the involved ingredients play.

kindly mail me

it would very helpfull to me

thanks a lot in advance


here is one :

NaCl 150mM (for osmolarity)
Tris, pH7.2 10 mM (for pH buffer)
triton X-100 1.0% (detergent)
deoxycholate 1% (an other detergent, you should not add it if you want to make some kinase activity analysis on your samples)
EDTA 5 mM (Ca chelator, will inhibit enzymes)
PMSF 1mM (protease inhibitor)
aprotinin, leupeptin, pepstatin 1 ug/mL of each (protease inhibitors)

if you want to analyse phosphorylation state, you should also add phosphatase inhibitors :
Na3VO4 (Na orthovanadate) 1mM
NaF 1 mM