CST, 74829S, Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying H3F3A (Lys27Met) mutation. Validated for Western Blotting,Immunofluorescence (Immunocytochemistry),Flow Cytometry (Fixed/Permeabilized). Available in 2 sizes. Highly specific and rigorously validated in-house, Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit Monoclonal Antibody (CST #74829) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunofluorescence (Immunocytochemistry): 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. For a carrier free (BSA and azide free) version of this product see product # 10376 . Protocol Available protocols: Western Blotting, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized) Specificity / Sensitivity Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit Monoclonal Antibody recognizes endogenous levels of K27M mutant histone H3.1, H3.2, and H3.3 proteins. This antibody may cross-react with wild-type histone H3.1, 3.2, and 3.3 when used at a high concentration. Careful titration of this antibody may be required to obtain optimal specificity. Species Reactivity: Human, Mouse Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to K27M mutant sequence of human histone H3.3 protein. Background Diffuse intrinsic pontine glioma (DIPG) is an aggressive brainstem astrocyte tumor arising mostly in children, leading to a long-term survival rate of less than 10%. Multiple whole-genome sequencing studies of DIPG patients identified commonly occurring mutations in the H3F3A gene encoding histone H3.3. One of these mutations, a lysine to methionine amino acid substitution (K27M), is found in up to 78% of DIPGs and 22% of non-brainstem pediatric gliomas (1-3). This mutation is associated with poor prognosis, with a mean survival time of 0.73 years for patients with the K27M mutation versus 4.6 years for patients without the mutation (1-3). Expression of the K27M mutant histone H3 is accompanied by a dramatic reduction in the levels of polycomb repressive complex 2 (PRC2)-mediated trimethylation of histone H3, changes in the distribution of PRC2 on the genome, and altered expression of genes associated with various cancer pathways (4-6). 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 Specification REACTIVITY: H M SENSITIVITY: Endogenous MW (kDa): 17 Source/Isotype: Rabbit IgG