Probing Chromatin Modifications in Response to ERK Signaling
Chromatin immunoprecipitation (ChIP) is a technique used to determine the association of proteins or histone modifications with chromatin regions in living cells or tissues, and is used extensively in the chromatin biology field to study transcriptional and epigenetic mechanisms. Increasing evidence points to an epigenetic coordination of signaling cascades, such as ERK, that regulate key processes in development and disease, revealing novel principles of gene regulation. Here we describe a detailed protocol for performing chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) for probing histone modifications regulated by ERK signaling in mouse ESCs.
Key wordsChromatin immunoprecipitation Histone modifications Polycomb ERK signaling Spike-in normalization ChIP-Western blot ChIP-qPCR
Work in the W.-W.T. lab is supported by research fundings from the Singapore National Research Foundation Fellowship as well as the Biomedical Research Council, Agency for Science, Technology and Research.