Hello all,
We're currently implementing a ChIP protocol on mice liver to analyze the binding of some TFs and the distribution of various histone modifications. One of These is H3K9me3. Our current results indicate that maybe the Ab we're using (abcam 8898) might not work. To clearly determine if our Problem is due to the Ab or the new protocol we would need a reliable positive control for H3K9me3 in mice.
It would be fantastic if anyone of you had fitting primers or an idea for a gene Region to design primers by ourselves. Thank you so much.
Perhaps a region around the telomerase TSS? Generally heavily methylated
How about the transcription start site of the telomerase gene? Should be heavily methylated.
Are you sure your primers work well. We bought anti-acetyl histone H3 from Millipore. At first we thought that Ab doesn't work but it turns out it's because of our primers. Also you can run a western blot to check your Ab.
Any of the imprinting control regions (eg H19 ICR) should be enriched for this mark and are often used as a control for this.
Alternatively, in the UCSC genome browser there are a number of tracks for histone modifications, including H3K9me3. Simply display these and you can search for an enriched site near your region of interest etc. UCSC has a number of data sets available, you can choose which best matches your cell type/experiment.
hello, have you solved your problem? Did you get the positive control primer?
check www.chipprimers.com if it's on there, the site is still under construction but I'm working on it! Feel free to send me any validated primers you have and I'll put them on there too.. alongside the PMID of your publication to cite.
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