Hello,
I hope I'm not doubling posts, I couldn't really find any threads on my exact issue.
OK. Let's assume that I want to ChIP genome wide for H3K4me1. I was thinking of best way of checking the specificity of binding of these kind of Ab. I had one idea: to do another ChIP in parallel for unmethylated version of the same epitope. Then my questions are:
1. Are there Abs available for methylated and unmethylated histones based on the very same peptide?
2. If there are, can I assume that K4me1 occurs at both H3 in a particular nucleosome? In other words, if there is enrichment for K4me1, does it mean that I won't see a signal for an unmethylated form or it is very well possible that one H3 can be monomethylated for K4 and the other one not?
3. What are your suggestions for QC of histone mod Abs?
Thank you,
Greg
I hope I'm not doubling posts, I couldn't really find any threads on my exact issue.
OK. Let's assume that I want to ChIP genome wide for H3K4me1. I was thinking of best way of checking the specificity of binding of these kind of Ab. I had one idea: to do another ChIP in parallel for unmethylated version of the same epitope. Then my questions are:
1. Are there Abs available for methylated and unmethylated histones based on the very same peptide?
2. If there are, can I assume that K4me1 occurs at both H3 in a particular nucleosome? In other words, if there is enrichment for K4me1, does it mean that I won't see a signal for an unmethylated form or it is very well possible that one H3 can be monomethylated for K4 and the other one not?
3. What are your suggestions for QC of histone mod Abs?
Thank you,
Greg
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