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nonspecific bands in silver stain IPs

nonspecific bands in silver stain IPs - Immunoprecipitation Forum

nonspecific bands in silver stain IPs - IP or Immunoprecipitation Forum.


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Old 09-30-2009, 05:33 PM
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Default nonspecific bands in silver stain IPs



Hi,

I'm trying to precipitate my protein of interest from total cell lysate, along with interacting proteins, so that they are visible on silver stained gels. The problem is that I am getting a lot of nonspecific bands sticking to control IPs (beads alone or with nonrelated antibody).

I have tried preclearing, and using higher ultracentrifugation, which have improved the stain a bit, but still too many nonspecific bands. Any suggestions? I'm about to try washing with different salt concentrations (usually use lysis buffer with 1% triton and 150mM NaCl).
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