SimpleChIP® Plus Sonication Chromatin IP Kit #56383

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Once in solution, store 1M DTT at -20°C.	-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed. -For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration. -For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation.

Upstream tips
-Once in solution, store 1M DTT at -20°C.
Protocol tips
-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed. -For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration. -For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation.

Publication protocol

ChIP assay was performed by using a ChIP assay kit (Cell Signaling). Briefly, SW620 cells were crosslinked with formaldehyde then chromatin was fragmented by digestion with Micrococcal Nuclease and sonication. Anti-NFAT1 or rabbit anti-IgG antibodies were used for precipitation, and the immunoprecipitates were purified with Agarose Beads. The purified ChIP DNA was adapter ligated and PCR amplified according to the manufacturer’s instructions (Illumina, San Diego, CA, USA). High-throughput sequencing was performed on an Illumina Genome Analyzer and short reads were mapped to the human reference genome. Peaks were assigned to specific genes by proximity to the nearest transcription start site. The differential peak-associated genes were annotated and clustered according to the KEGG pathway and Reactome databases.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for SimpleChIP® Plus Sonication Chromatin IP Kit #56383 below.

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