ChIPAb+ EZH2; clone AC22

ChIPAb+ EZH2; clone AC22

Stock Code: 3586465
Manufacturer Part No: 17-662
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Analysis Note


ControlIncludes negative control mouse IgG antibody and primers specific for of human MYT-1 promoter region.


Application


Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 µg of either a normal mouse IgG or Anti-EZH2, clone AC22 antibody and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of EZH2-associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and MYT-1 promoter (positive) primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Immunoprecipitation Analysis:
Representative lot data.
HeLa nuclear cell extract was Immunoprecipitated with either anti-EZH2, clone AC22 or mouse IgG and then resolved by electrophoresis, transferred to PVDF and probed with Cat. # 05-1319 anti-EZH2, clone BD43 (1:1000 dilution).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. # AP124P) and a chemiluminescence detection system.
(Please see figures).
Lane 1: Hela nuclear extract immunoprecipitate with anti-EZH2, clone AC22.
Lane 2: Hela nuclear extract immunoprecipitate with IgG.


EZH2, clone AC22, ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of EZH2.


Research CategoryEpigenetics & Nuclear FunctionApoptosis & Cancer


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


EZH2 is a polycomb group (PcG) protein that is a catalytic subunit of the PRC2/EED-EZH2 complex, which methylates ′Lys-9′ (H3K9me) and ′Lys-27′ (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate ′Lys-27′ of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes

Quality Level100
ManufacturerSIGMA-ALDRICH

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