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Lysis buffer for Immunoprecipitation


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#1 mtrnbh

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Posted 09 October 2009 - 10:21 AM

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

#2 Dr Teeth

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Posted 09 October 2009 - 10:41 AM

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


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 mtrnbh

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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.




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