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  • jdub
    Junior Member
    • Nov 2010
    • 8
    #1

    Non-specific binding of control antibodies for ChIP-seq

    I am having trouble with a lot of non-specific binding with both my negative and positive control antibodies. I am using an anti-RNA Pol II positive control and rabbit IgG negative control and using primers for GAPDH and ZNF333. I get equivalent levels of amplicons at both loci with both antibodies. The concentrations used are what was recommended for IP...25 ug/mL for anti-RNA Pol II and 100 ug/mL for IgG.

    Does anyone have any suggestions on where my ChIP might be going wrong?
    Tags: None
    • jdub
      Junior Member
      • Nov 2010
      • 8
      #2
      By the way, I have tried two different cell lines and gotten the same result with each.

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