Washing buffer and use of Protease inhibitors - (Oct/14/2011 )
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.
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.