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Allele-specific iChIP-SILAC analysis to identify proteins binding to the p16INK4A gene

ABSTRACT: We used insertional ChIP (iChIP) based approach to identify proteins that bind to the p16INK4A gene in the human cell line HCT116. To perform iChIP, we inserted an 8-repeat of the LexA-binding element (LexA-BE) in the p16INK4A promoter region in HCT116. The DNA-binding domain and dimerization domain of the LexA protein fused with the 3xFLAG tag and a nuclear localization signal (NLS) was expressed in the LexA-BE-knock-in cells (#109-2 and #113-5). The resultant cells (#109-2-1 and #113-5-1) were subjected to stable isotope labeling with amino acids in cell culture (SILAC), immunoprecipitated with an anti-FLAG antibody, and subjected to LC-MS/MS analysis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Toshitsugu Fujita

LAB HEAD: Hodaka Fujii

PROVIDER: PXD045843 | Pride | 2024-04-18

REPOSITORIES: Pride

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Identification of immunoglobulin switch (S) region binding proteins by insertional ChIP (iChIP)

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