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Washing buffer and use of Protease inhibitors


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#1 Marvilla

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Posted 14 October 2011 - 07:50 AM

Hi, I have a question regarding the washing step in ChIP

The buffer for washing the immunoprecipitated DNA-bead complex should contain protease inhibitors? I guess that at the first wash there's still a significant amount of proteases from cell lysate but at the following washings I believe that there's no need to use protease inhibitors or use a lower amount. Please tell me how you do it. Thanks.

#2 chabraha

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Posted 17 October 2011 - 09:36 AM

I never add protease inhibitors to any of the wash buffers and I think it doesn't matter since my wash steps are ~5min, also some of the protease inhibitors you should be using irreversibly inhibit proteases.
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