Protocol Online logo
Top : New Forum Archives (2009-): : Biochemistry

which is the best lysis buffer for whole cell extracts? - (Apr/15/2010 )

hi,

I wanna do a whole cell extract from HEK cells to detect telomerase protein by western blot.
So far I used WB only for cytoplasmic protein so the lysis buffer that I used (10 mM Tris/HCl, 10 mM EDTA, 100 mM NaCl, 0,5 NP40, 0,5% Desoxycholat) seems not to disrupt the nuclear membrane (this is what I read from the literature...) .
I need a lysis buffer that disrupt the nuclear membrane. could someone help me with a protocol??

thx a lot,
georgeta

-georgeta-

Try RIPA buffer.

-bob1-