Application

Anti-Histone H3.3 K9M, Cat. No. ABE579, is a highly specific rabbit polyclonal antibody that setects Histone H3.3 mutant K9M and has been tested for use in Western Blotting.

Western Blotting Analysis: A representative lot detected Histone H3.3 K9M in immunoblots of affinity-purified heterotypic mononucleosomes from lentivirus-transduced 293T cells expressing various epitope-tagged H3.3 transgenes (Jayaram, H., et. al. (2016). Proc Natl Acad Sci USA. 113(22):6182-7).

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Histone H3.3 K9M in whole cell extract of mouse embryonic fibroblasts (MEF) or mouse embryonic stem cells (mESCs) expressing various H3.3 constructs (Courtesy of Dr. Peter Lewis, Rockefeller University).

Research CategoryEpigenetics & Nuclear Function

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 H3.3 (UniProt: P84243) is encoded by the H3F3A (also known as H3.3A, H3F3, PP781, H3F3B, H3.3B) gene (Gene ID: 3020) in human. It is a highly conserved variant form of Histone H3, which replaces conventional H3 in a wide range of nucleosomes in active genes. Histone H 3.3 constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. H3.3 is associated with actively expressed genes in both animals and plants. It is predominantly enriched near transcription end sites (TES) of genes and positively associated with transcription. Histone H3 contains a main globular domain and a long N-terminal tail and is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. For example, acetylation on Lys10 (H3K9ac; initial methionine removed) impairs methylation at Arg9 (H3R8me2s) and acetylation on Lys19 (H3K18ac) and Lys24 (H3K24ac) favors methylation at Arg18 (H3R17me). Phosphorylation at Ser11 (H3S10ph) by Aurora kinase B is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. Phosphorylation at Ser11 by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-Fos and c-Jun. Mutations in Histone H3.3 have been implicated in a high proportion of malignant pediatric brain cancers. The mutant H3.3 histone disrupts epigenetic post-translational modifications near genes involved in cancer processes and in brain function. Lysine to methionine mutations in genes encoding histone H3 are believed to be responsible for subset of pediatric brain and bone cancers. K9M residues that can occupy the active site cavity of G9a methyltransferase and act as competitive inhibitor. G9a methyltransferase has been shown to be important in heterochromatin maintenance and transcriptional repression. K9M can lead to a reduction in global levels of H3K9me3 in cells. (Ref.: Jayaram, H et al. (2016). Proc. Natl. Acad. Sci. USA 113(22); 6182-6187).

Immunogen

KLH-conjugated linear peptide corresponding to 9 amino acids from the N-terminal region of human histone H3.3 with K9M mutation.

Epitope: N-terminus

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Affinity Purified

Format: Purified

Quality

Evaluated by Western Blotting in HEK293 cells transfected with Histone H3.3 WT and Histone H3.3 K9M mutant.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Histone H3.3 K9M in HEK293 cells transfected with H3.3 K9M mutant. It did not detect wild-type Histone H3.3.

Specificity

This rabbit polyclonal antibody detects K9M mutant form of histone H3.3 and does not react with wild-type H3.3.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~19 kDa observed; 15.33 kDa calculated. Uncharacterized bands may be observed in some lysate(s).