05-1242-S

Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4, Trial Size

clone 6F12-H4, from mouse

• Synonym(s): H3K9me3, Histone H3 (trimethyl K9), H3 histone family, member T, Histone 3, H3, Histone cluster 3, H3
• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Properties

• biological source: mouse
• Quality Level: 100
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 6F12-H4, monoclonal
• species reactivity: mouse, human
• technique(s): ChIP: suitable (ChIP-seq); dot blot: suitable; immunofluorescence: suitable; inhibition assay: suitable (peptide); western blot: suitable
• isotype: IgG1κ
• NCBI accession no.: NM_003493
• UniProt accession no.: Q16695
• shipped in: wet ice
• target post-translational modification: trimethylation (Lys9)
• Gene Information: human ... H3C1(8350) ; mouse ... H3C1(360198)

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.

Specificity

Based on sequence homology, broad species cross-reactivity is expected with all mammals, Drosophila, Xenopus, and Arabidopsis.

Recognizes trimethyl-histone H3 (Lys9), Mr 17 kDa. Cross-reacts with dimethyl histone H3(Lys9) at higher concentrations.

Immunogen

Epitope: Trimethyl Lys9

KLH-conjugated, branched synthetic peptide containing the sequence (QTARme3K- STGGKA)2-KC, in which me3K corresponds to trimethyl lysine 9 of human histone H3. Data suggests this antibody may require the presence of the glutamine residue in position 5 for binding.

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.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

Use Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 (mouse monoclonal antibody), Trial Size / Pack validated in ChIP, PIA, IF, DB, ChIP-seq, WB to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).

Quality

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.

Target description

~17 kDa

Physical form

Format: Purified

Purified

Purified antibody supplied at 1 mg/ml in 0.1 M Tris-glycine, pH 7.4, 150 mM NaCl, 0.05% NaN3.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid freezing, which can damage IgG and affect product performance.

Analysis Note

Control
HeLa Acid Extract

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.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable