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which is the best lysis buffer for whole cell extracts?


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#1 georgeta

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Posted 15 April 2010 - 01:44 AM

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

#2 bob1

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Posted 15 April 2010 - 04:54 PM

Try RIPA buffer.




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