ChIPAb+ ère

Non disponible en dehors du Royaume-Uni et de l'Irlande

Analysis Note

ControlIncludes control primers specific for human TFF1 (pS2) promoter.

Application

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

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 CategoryEpigenetics & Nuclear Function

Research Sub CategoryChromatin Biology

Disclaimer

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.

General description

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.

Immunogen

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

Legal Information

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

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

Packaging

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

Physical form

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

Quality

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.

Specificity

Estrogen Receptor α

Storage and Stability

1 year at -20°C from date of shipment

Target description

66 kDa