Description
General description
Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely via acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α/β. The enzymes responsible for H3K9me3 formation are SUV39H1 and SUV39H2.
~17 kDa
Application
Chromatin Immunoprecipitation (ChIP):
Representative data from a previous lot. Sonicated 3T3 L1 chromatin was subjected to chromatin immunoprecipitation using anti- trimethyl-histone H3 (Lys9) and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of trimethylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using primers flanking the p16 promoter.
Peptide Inhibition Analysis:
Peptide blocking assay demonstrates distinct preference of the antibody for the trimethyl form vs. the dimethyl form.
Chromatin Immunoprecipitation (ChIP):
ChIP analysis of known chromosomal Suv39h targets (H3K9me3 in major satellites, mouseES cells).
Dot Blot Analysis:
Dot-blot analysis demonstrating specificity of anti-H3K9me3, clone 6F12-H4 for trimethyl Lys9 of histone H3.
Use Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 (mouse monoclonal antibody) validated in ChIP, PIA, IF, DB, ChIP-seq, WB to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).
Biochem/physiol Actions
Based on sequence homology, broad species cross-reactivity is expected with all mammals, Drosophila, Xenopus, and Arabidopsis.
Physical form
Format: Purified
Analysis Note
Routinely evaluated by Western Blot on HeLa acid extracts.
Western Blot Analysis: A 0.5 – 5 μg dilution of this lot detected trimethyl histone H3 (Lys9) in HeLa acid extracts.
