ChIP Assay for Protein-DNA Interactions

A ChIP assay was performed using a ChIP Assay Kit (Millipore) according to the manufacturer’s instructions. Briefly, LN-229 cells were cross-linked and lysed, and DNA was sheared into 200–800 bp fragments using sonication. Precleared chromatin was immunoprecipitated with a Flag antibody (Santa Cruz), and DNA was isolated after reverse cross-linking for quantitative real-time PCR (qRT-PCR). The relevant primer sequences are presented in Table S3 in the Supplemental Materials. ChIP analyses were then performed as previously described^{33 (link)}.

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Deng Q., Hou J., Feng L., Lv A., Ke X., Liang H., Wang F., Zhang K., Chen K, & Cui H. (2018). PHF19 promotes the proliferation, migration, and chemosensitivity of glioblastoma to doxorubicin through modulation of the SIAH1/β–catenin axis. Cell Death & Disease, 9(11), 1049.

Publication 2018

ChIP Assay Kit Flag antibody

Antibody Cells Chip Chip assay Chromatin Primer Quantitative real time pcr

Corresponding Organization :

Other organizations : Southwest University, Daping Hospital, Army Medical University

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Variable analysis

independent variables

Antibody used for immunoprecipitation (Flag antibody)

dependent variables

DNA fragments enriched in the immunoprecipitation

control variables

Cell line used (LN-229 cells)
Cross-linking and lysis procedure
Chromatin shearing method (sonication to 200-800 bp fragments)
Quantification method (quantitative real-time PCR)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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