ChIPAb+ Dimethyl-Histone H3 (Lys9)

Stock Code: 3586458
Manufacturer Part No: 17-648

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Descriptions

Analysis Note

ControlIncluded negative control antibody normal rabbit serum and control primers specific for human β-globin promoter.

Application

Dimethyl-Histone H3 (Lys9) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K9Me2.

Research CategoryEpigenetics & Nuclear Function

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 10⁶ cell equivalents) was subjected to chromatin Immunoprecipitation using 4 µL of either a normal rabbit antiserum or Antidimethyl-Histone H3 (Lys9) serum and the Magna ChIP A (Cat. #17-610) Kit (Please see figures). Successful Immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using β-globin ChIP Primers flanking the human β-globin promoter or primers amplifying the promoter of human GAPDH, which is transcriptionally inactive in HeLa cells. Percent Input relative to standard curves for each qPCR primer set are shown.
Please refer to the EZ-Magna A ChIP™ (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western blot analysis and peptide inhibition:
HeLa Acid extract were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-dimethyl-Histone H3 (Lys9) (1:500, Lane 1) or preincubated with 0.4 µM Histone H3 peptide with following modifications:
Lane 2: Linear non-modified
Lane 3: Branched non-modified
Lane 4: Branched trimethyl
Lane 5: Linear trimethyl
Lane 6: Branched dimethyl
Lane 7: Linear dimethyl
Lane 8: Branched monomethyl
Lane 9: Linear monomethyl
Proteins were visualized using a goat anti–rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.

Research Sub CategoryChromatin Biology

Components

Anti-Dimethyl-Histone H3 (Lys9) (rabbit serum), 1 vialNegative ChIP Control serum, 1 vialChIP Primers β-globin , 1 vial

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

The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 9 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SuvH39H1 or G9a. This methylated lysine can be demethylated by histone demethylases as JMJD1A, LSD1 or JMJD2C. Methylation of this residue is mainly associated with transcriptional repression.

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context