CTCF (D31H2) XP® Rabbit mAb #3418

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-It is highly critical that the chromatin is of appropriate size and concentration.	-For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. -It is important to keep the tissue cold to avoid protein degradation. -Use fresh formaldehyde that is not past the manufacturer's expiration date.	-Once in solution, store 1M DTT at -20°C.

Upstream tips
-It is highly critical that the chromatin is of appropriate size and concentration.
Protocol tips
-For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. -It is important to keep the tissue cold to avoid protein degradation. -Use fresh formaldehyde that is not past the manufacturer's expiration date.
Downstream tips
-Once in solution, store 1M DTT at -20°C.

Publication protocol

Chromatin was prepared from MCF-7 cells by fixing the cells in 37% formaldehyde for 15 min, followed by quenching with glycine (2.5 M) for 10 min. Sonication of cross-linked genomic DNA was performed using Sonics Vibra CellTM to obtain 500~1,000 bp DNA fragments. ChIP was performed using antibodies against CTCF (#3418; Cell Signaling), PolII (sc-56767; Santa Cruz Biotechnology, CA, USA), H3K4me3 (#1012; Abcam, Cambridge, UK), H3K27me3 (ab6002; Abcam), and IgG (Santa Cruz, sc-2027).

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for CTCF (D31H2) XP® Rabbit mAb #3418 below.

Download manufacturer protocol

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