SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003

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

T47D cells were treated with VC (ethanol), R5020, TPA, or R5020+TPA for 30 minutes. Chromatin immunoprecipitation (ChIP) was performed with the SimpleChIP Enzymatic Chromatin IP Kit (Cell Signaling, Danvers, MA) according to the manufacturer’s instructions. Rabbit IgG (Cell Signaling), p300 (Santa Cruz Biotechnology; catalog no. sc-585; RRID: AB_2231120) (33), and PR (Santa Cruz Biotechnology; catalog no. sc-7208; RRID: AB_2164331) (29) antibodies were used for ChIP. Immunoprecipitated DNA was extracted with the QIAquick PCR purification kit (Qiagen), and DNA was subjected to real-time PCR analysis. Primer sequences were obtained from a previously published study (34). Enrichment was calculated as a percentage of total input DNA. All experiments were performed in triplicate.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 below.

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