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

HSC-T6 cells stimulated with TGF-β1 (10 ng/mL), and corresponding control cells were cultured to ~80-90 % confluency. Subsequently, ChIP assay was performed using the SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 (Cell Signaling Technology, USA) according to the manufacturer's instructions. The following antibodies were used to immuno-precipitate crosslinked protein-DNA complexes: rabbit anti-DNMT1, rabbit anti-DNMT3a, rabbit anti-DNMT3b and normal rabbit IgG. The immunoprecipitated DNA was purified for PCR analyses with primers specific for the putative binding sites within the promoter of RCAN1.4.

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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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