Histone H3 (K36M Mutant Specific) (E8N1L) Rabbit mAb #98399

Filter:

WB
IF
ChIP

Supporting Data

REACTIVITY	H M
SENSITIVITY	Endogenous
MW (kDa)	17
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IF-Immunofluorescence
ChIP-Chromatin Immunoprecipitation

Species Cross-Reactivity Key:

H-Human
M-Mouse

Product Information

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 x 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

Application	Dilution
Western Blotting	1:1000
Immunofluorescence (Immunocytochemistry)	1:1600 - 1:6400
Chromatin IP	1:50

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

Specificity / Sensitivity

Histone H3 (K36M Mutant Specific) (E8N1L) Rabbit mAb recognizes endogenous levels of K36M mutant histone H3.1, H3.2, and H3.3 proteins. The antibody may show slight cross-reactivity with wild-type histone H3.1, 3.2, or 3.3 when used at a high concentration. Careful titration of this antibody may be required to obtain optimal specificity. This antibody may detect a band of unknown origin at 50 kDa.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to K36M mutant sequence of human histone H3.3 protein.

Background

Chondroblastoma is a rare type of benign tumor that is found at the rounded ends of the long bones in the arms and legs. More than 90% of chondroblastomas have been found to contain a heterozygous mutation in the H3F3A gene encoding the histone variant H3.3 (1). This mutation, a lysine to methionine amino acid substitution in codon 36 (K36M), inhibits at least two histone H3 lysine 36 methyltransferases, WHSC1 (MMSET) and SETD2, resulting in reduced global levels of histone H3 lysine 36 methylation (1). Chondrocytes containing the histone H3 K36M mutation exhibit several hallmarks of cancer cells, including increased ability to form colonies, resistance to apoptosis, and defects in differentiation. Reduction of global methylation levels in chondrocytes, resulting from the K36M mutation, contributes to tumorigenesis by altering the expression of cancer-associated genes. The histone H3 K36M mutation is also found to promote sarcomagenesis by impairing the differentiation of mesenchymal progenitor cells, resulting in undifferentiated sarcomas (2). The K36M mutation alters the histone methylation landscape, resulting in a genome-wide gain in histone H3 lysine 27 methylation, redistribution of polycomb repressive complex 1, and derepression of its target genes known to block mesenchymal differentiation. Finally, the histone H3 K36M mutation is also found in 13% of HPV-negative head and neck squamous cell carcinomas, again contributing to tumorigenesis by altering global methylation levels of histone H3 lysine 36 (3).

Database Links

UniProt ID: P84243

Entrez-Gene Id: 3020

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.

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