ChIP Assay for SREBP1 Binding

ChIP was performed as described previously^{31 (link)}using SREBP1 antibody (Santa Cruz, sc-367x). Briefly, 5 μg rabbit IgG (Santa Cruz) or SREBP1 antibody was incubated with Protein A Dynabead magnetic beads (Invitrogen) for 4 h, followed by extensive washing to remove unbound antibody. Antibody beads were then added to the chromatin and incubated overnight. The ChIP primers were purchased from Qiagen (EpiTect ChIP PCR assay) and used for qPCR analysis: BCAT2 (GPH1020848(+)03A).

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Dey P., Baddour J., Muller F., Wu C.C., Wang H., Liao W.T., Lan Z., Chen A., Gutschner T., Kang Y., Fleming J., Satani N., Zhao D., Achreja A., Yang L., Lee J., Chang E., Genovese G., Viale A., Ying H., Draetta G., Maitra A., Wang Y.A., Nagrath D, & DePinho R.A. (2017). Genomic deletion of malic enzyme 2 confers collateral lethality in pancreatic cancer. Nature, 542(7639), 119-123.

Publication 2017

SREBP1 antibody Protein A Dynabead magnetic beads EpiTect ChIP PCR assay

Antibody Chip Chip assay Chromatin Primers Protein a Rabbit Srebp1

Corresponding Organization :

Other organizations : The University of Texas MD Anderson Cancer Center, Rice University

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Variable analysis

independent variables

SREBP1 antibody (Santa Cruz, sc-367x)
Rabbit IgG (Santa Cruz)

dependent variables

BCAT2 gene expression (measured by qPCR)

control variables

Protein A Dynabead magnetic beads (Invitrogen)
ChIP primers (Qiagen, EpiTect ChIP PCR assay)

controls

Positive control: SREBP1 antibody
Negative control: Rabbit IgG

Annotations

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