ChIPAb+ Estrogen Receptor α - ChIP Validated Antibody and Primer Set

Every lot of the ChIPAb+ line of antibodies is individually validated for chromatin precipitation, in order to guarantee successful ChIP assays every time. Each antibody includes a control primer set for performance confirmation. ERα antibody is functionally validated in the precipitation of ERα associated chromatin.
The qPCR primers included flank the ERα binding site in human pS2 promoter.

Estrogen Receptor α

The ERα antibody is made against aa 120-170 of bovine estrogen α.

Chromatin Immunoprecipitation:
Sonicated chromatin, prepared from 3x10⁶ MCF7 cells that are either estrogen starved or b-estradiol treated (100 nM, 45 min.) was subjected to chromatin
immunoprecipitation using 4 mL anti-ERa and the Magna ChIP^® G (Cat.# 17-611) kit Standard Protocol. Successful enrichment of ER associated DNA fragments was verified by qPCR using ChIP Primers TFF1 (pS2) flanking the human TFF1 promoter that contains an ER binding site (Please see figures).
Please refer to the EZ-Magna ChIP G (Cat.# 17-409) or EZ-ChIP (Cat.# 17-371) kit protocols for experimental details.

Western Blot Analysis:
MCF7cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-ERa (1:1000 dilution). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

This ChIPAb+ Estrogen Receptor α -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

25 assays per kit. ~4 μL per chromatin immunoprecipitation.

Chromatin immunoprecipitation:
Sonicated chromatin prepared from 3x10⁶ MCF7 cells that were treated with b-estradiol (100 nM, 45 min.) was subjected to chromatin immunoprecipitation using 4 mL anti-ERα and beads only control.
Successful enrichment of ERα associated DNA fragments was verified by qPCR using ChIP Primers TFF1 (pS2) flanking the human TFF1 promoter that contains an ER binding site (Please see figures).
Please refer to the EZ-Magna ChIP G (Cat.# 17- 409) or EZ-ChIP (Cat.# 17-371) kit protocols for experimental details.

66 kDa

Anti-ERα (mouse ascites). 1 vial containing 100 mL ascites. Store at -20°C. The ERα antibody is made against aa120-170 of bovine estrogen a. It can recognize human and mouse ERα.

ChIP primers TFF1 (pS2). 1 vial containing 75 mL of 5 μM of each control primer specific for human TFF1 (pS2) promoter. Store at -20°C.
FOR: CCG GCC ATC TCT CAC TAT GAA
REV: CCT TCC CGC CAG GGT AAA TAC

1 year at -20°C from date of shipment

Control
Includes control primers specific for human TFF1 (pS2) promoter.

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.