Analysis Note

ControlAcid extracted proteins from HeLa cells.

Application

Peptide Inhibition Analysis:
10 µM of a Histone H3 peptide containing trimethyl-lysine 9 abolished detection of Histone H3 by anti-trimethyl Histone H3 (Lys9) in western blot analysis of HeLa acid extracts (Figure B, lane 4). Slight signal reduction was observed with 10 µM of a Histone H3 peptide containing dimethyl-lysine 9.

Peptide Dot Blot Analysis:
A 1:2000 dilution of a previous lot was incubated with serial dilutions of Histone H3 peptides containing various modifications spotted onto PVDF. Slight cross-reactivity with Histone H3 peptide containing dimethyl-lysine 9 was detected.

Beadlyte™ Histone Peptide Assay:
1:2000-1:6000 dilutions of a previous lot were incubated with Histone H3 peptides containing various modifications conjugated to Luminex™ microspheres. Slight cross-reactivity with Histone H3 peptides containing dimethyl-lysine 9 and trimethyl-lysine 27 was detected.

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

Anti-trimethyl-Histone H3 (Lys9) Antibody is a Rabbit Polyclonal Antibody for detection of trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9) & has been validated in DB, PIA & WB.

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

Histone H3 is a core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

Immunogen

Epitope: Trimethyl Lys9

BSA-conjugated, synthetic peptide containing the sequence R30;AR[me3K]SR30; in which me3K corresponds to trimethyl-lysine 9 of human Histone H3.

Legal Information

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

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Rabbit polyclonal antiserum IgG in buffer containing 0.05% sodium azide before the addition of 30% glycerol.

Immunodepleted Serum

Quality

Evaluated by western blot in acid-extracted proteins from HeLa cells.

Western Blot Analysis:
1:500-1:5000 dilution of this antibody detected methylated Histone H3 (Lys9) in acid-extracted proteins from HeLa cells. The antibody did not detect unmethylated recombinant Histone H3 (Catalog # 14-411).

Specificity

A broad species cross-reactivity is expected.

Recognizes Histone H3 containing trimethyl-lysine 9 and, to a lesser extent, dimethyl-lysine 9. MW ~17 kDa.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Target description

~17 kDa