Application

Research CategoryEpigenetics & Nuclear Function

Anti-Histone H3.3 K36I Mutant, Cat. No. ABE1450, is a highly specific rabbit polyclonal antibody that detects Histone H3.3 Mutant (K36I) and has been tested for use in Western Blotting.

Western Blotting Analysis: A 1:1,000 dilution from a representative lot specifically detected Histone H3.3 K36I Mutant in HEK293 cells transfected with H3.3 K36I mutant, but not the wild-type or K36M mutant (Courtesy of Dr. Peter Lewis, Rockefeller University).

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 H3.3 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. K36I mutations in Histone H3 have been reported in pediatric undifferentiated sarcoma tumors. K36I mutation is shown to inhibit H3K36 methylation, which can lead to a genome-wide gain in H3K27 methylation that leads to redistribution of polycomb repressive complex 1 and de-repression of its target genes to block mesenchymal differentiation.

Immunogen

KLH-conjugated linear peptide corresponding to 8 amino acids from the N-terminal region of human histone H3.3 with K36I mutation.

Epitope: unknown

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 expressing Histone H3.3 K36I Mutant.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Histone H3.3 K36I Mutant in HEK293 cells expressing Histone H3.3 K36I Mutant.

Specificity

This rabbit polyclonal antibody detects K36I 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).