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

Chromatin immunoprecipitation (.ChIP) assay was carried out using SimpleChIP® Enzymatic Chromatin IP Kit (Cell Signaling, Danvers, MA, US) in accordance to the manufacturer’s instructions. Chromatin fragments of HCC cells were immunoprecipitated with 5 μg anti-PHF8 or IgG antibody (Abcam) and then subjected to qRT-PCR using SYBR Premix Ex Taq kit (Takara) performed on an ABI Prism 7500 Real-Time System. Methods for detection and calculation of target amplification were according to previous descriptions [28]: fold enrichment = 2− (ΔCT expt - ΔCT control) where ΔCT = CT anti-PHF8-IP–CT IgG-IP, expt = target region, and control = negative control region which was from an intragenic genome. Sequence covering the region of − 2.5 kb upstream and 1.0 kb downstream of TSS was selected for designing primers.

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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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