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High background in negative loci RNA Pol II Chip


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#1 trungbiotech

trungbiotech

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Posted 27 November 2014 - 10:17 PM

Hi everyone,

Can you help me with this?

I am doing Chip using the  EZ-Magna ChIP™ HiSens Chromatin Immunoprecipitation Kit from Millipore. I have done Chip on MeCP2(methylated binding protein) and it worked perfectly etc I had good enrichment from the positive control region versus the negative control regions and good enrichment against the IgG as well

The problem is when I moved on to do the RNA Pol II Ser2P Chip and I am using the antibody from abcam (ab5095) I always have enrichment in the intergenic regions which RNA Pol II-ser2P not supposed to bind to. This enrichment is even higher than my positive controls (gene bodies Gapdh and Actb)

Has anyone experienced the same thing?






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