Protocol detail

Chromatin Immunoprecipitation of SETDB2

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MKN74 and MKN45 cells were transfected with SETDB2 siRNA or negative control siRNA for 48 hours. ChIP assays were performed using the ChIP-IT Express Kit (Active Motif) according to the manufacturer's instructions [9 (link)]. AGS cells were transfected with the SETDB2/pcDNA3 or empty/pcDNA3 expression vector. Antibodies used were anti-H3 (Active Motif), anti-H3K9me3 (Active Motif), Normal Rabbit IgG (No. 2729, Cell Signaling Technology, Danvers, MA) and anti-SETDB2 (No. 5517, abcam).

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Nishikawaji T., Akiyama Y., Shimada S., Kojima K., Kawano T., Eishi Y., Yuasa Y, & Tanaka S. (2016). Oncogenic roles of the SETDB2 histone methyltransferase in gastric cancer. Oncotarget, 7(41), 67251-67265.

Publication 2016

ChIP-IT Express Kit anti-H3 anti-H3K9me3

Antibodies Cells Chip Chip assays Rabbit Sirna Vector

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Variable analysis

independent variables

SETDB2 siRNA
Negative control siRNA
SETDB2/pcDNA3 expression vector
Empty/pcDNA3 expression vector

dependent variables

H3 occupancy
H3K9me3 occupancy
SETDB2 occupancy

control variables

Cell lines: MKN74, MKN45, AGS

controls

Positive control: Anti-H3 antibody
Negative control: Normal Rabbit IgG

Annotations

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