Hi all,
Can I use lysis buffer containing detergents for cell lysis prior to IP, I mean does detergent interfere with antibody binding? If so could someone who ha done IP recommend a suitable lysis buffer recipe?
Thanks in advance
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2 replies to this topic
#2
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Posted 09 October 2009 - 10:41 AM
mtrnbh, on Oct 9 2009, 02:21 PM, said:
Hi all,
Can I use lysis buffer containing detergents for cell lysis prior to IP, I mean does detergent interfere with antibody binding? If so could someone who ha done IP recommend a suitable lysis buffer recipe?
Thanks in advance
Can I use lysis buffer containing detergents for cell lysis prior to IP, I mean does detergent interfere with antibody binding? If so could someone who ha done IP recommend a suitable lysis buffer recipe?
Thanks in advance
The amount of detergent to use will depend on your antibody and other assay conditions. I have used 0.1 to 5% Triton X-100.
My cell lysis buffer is:
50 mM Tris-HCl, pH 7.4
150 mM NaCl
1 mM EDTA
1 mM EGTA
5 mM MgCl2
0.5% Triton x-100
0.2 mg/ml BSA
Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley
#3
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Posted 09 October 2009 - 09:49 PM
Thank you very much for your reply, I'll try your recipe; I hope it will work.
