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This kit is specially designed for entry level researchers to start performing the ChIP assay. In my case, I needed to test the interaction of various intracellular signaling molecules and determine the downstream gene target. This kit contain pre-made reagents to perform the ChIP assay using protein extracts from cells or tissues through the collection of the immunoprecipitated protein and DNA fragments which are then ready to analyze via Western blot or RT-PCR. The protocol is very simple and suggests which controls may be required for the assay.
ChIP assay
Mouse C2C 12 cell line
I crosslinked treated cells with 1% paraformaldehyde for 10 minutes. I then prepared cell lysates from 1 million cells with the indicated buffer from the kit. The cell lysate was sonicated and diluted and IP’ed with the recommended ChIP grade antibody (also from EMD Millipore), for an overnight incubation. The next day, the chromatin-protein complexes were washed with a series of buffers provided by the kit. I either directly performed Western blotting of the IP sample or amplified the gene promoter region after reverse-crosslink by real-time RT-PCR.
To achieve better results from the CHIP assay kit, it is very important to pre-qualify the system. For example, it is important to determine the optimal cell concentration to start with, and to determine the time and intensity of crosslinking and sonication. Usage of a ChIP grade antibody for the IP is also important. And a good positive and negative control of each step is also very important for the quantification of the results.
Overall, the CHIP assay kit performed well for my experiments. The protocol is simple and gives results showing protein interaction and promoter amplification.
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Product Name: Chromatin Immunoprecipitation (ChIP) Assay KitSupplier Page