Description
Product Usage Information
Western Blotting: 1:1000
Immunoprecipitation: 1:50
Immunofluorescence (Immunocytochemistry): 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized): 1:400 - 1:1600
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized)
Specificity / Sensitivity
Histone H3 (K9M Mutant Specific) (E2E9L) Rabbit Monoclonal Antibody recognizes endogenous levels of K9M mutant histone H3.1, H3.2, and H3.3 proteins. This antibody does not cross-react with wild-type histone H3.1, H3.2, or H3.3.
Species Reactivity: Human, Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to K9M mutant sequence of human histone H3.3 protein.
Background
Multiple exome sequencing analyses have uncovered a high frequency of histone H3 driver mutations in a number of different cancers, including diffuse intrinsic pontine glioma (DIPG), chondroblastoma, sarcomas, and HPV-negative head and neck squamous cell carcinoma. Previous studies have shown that lysine to methionine histone mutations in these cancers act as potent inhibitors of their respective lysine methyltransferases, resulting in gross alterations to the histone methylation landscape and deregulation of gene expression. In DIPG for example, the histone H3 K27M mutation is accompanied by a dramatic reduction in the levels of polycomb repressive complex 2 (PRC2)-mediated tri-methylation of histone H3 lysine 27, changes in the distribution of PRC2 on the genome, and altered expression of genes associated with various cancer pathways (1-3). In chondrocytomas, the histone H3 K36M mutation functions to inhibit the WHSC1 (MMSET) and SETD2 histone methyltransferases, resulting in a reduction in the levels of histone H3 lysine 36 tri-methylation and deregulation of a number of cancer-associated genes (4). Similar to the H3K27M and H3K36M mutations, the histone H3 K9M mutation has been shown to inhibit the H3K9-directed histone methyltransferase G9a, resulting in reduced levels of histone H3 lysine 9 trimethylation (5). Given the widespread role of G9a in the regulation of gene expression, it is likely that this K9M mutation also plays a role in cancer.
Alternate Names
H3 histone family member 3A; H3 histone, family 3A; H3 histone, family 3B (H3.3B); H3-3A; H3-3B; H3.3 histone A; H3.3A; H3.3B; H33; H3F3; H3F3A; H3F3B; Histone H3.3
