Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

17

SOURCE:

Rabbit

UniProt ID:

#P84243

Entrez-Gene Id:

3020

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Histone H3 (K9M Mutant Specific) Antibody recognizes endogenous levels of K9M mutant histone H3.1, H3.2, and H3.3 proteins. The antibody does not cross-react with wild-type histone H3.1, 3.2, or 3.3.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to K9M mutant sequence of human histone H3.3 protein. Antibodies are purified by protein A and peptide affinity chromatography.

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.

  1. Chan, K.M. et al. (2013) Genes Dev 27, 985-90.
  2. Lewis, P.W. et al. (2013) Science 340, 857-61.
  3. Piunti, A. et al. (2017) Nat Med 23, 493-500.
  4. Fang, D. et al. (2016) Science 352, 1344-8.
  5. Jayaram, H. et al. (2016) Proc Natl Acad Sci U S A 113, 6182-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 1
#78087

Histone H3 (K9M Mutant Specific) Antibody

Western Blotting Image 1: Histone H3 (K9M Mutant Specific) Antibody Expand Image
使用 Histone H3 (K9M Mutant Specific) Antibody(上图)和 Histone H3 (D1H2) XP® Rabbit mAb #4499(下图)对野生型 mES 细胞 (mES/H3.3 WT) 和 K9M 突变型组蛋白 H3.3 敲入 mES 细胞 (mES/H3.3 K9M) 的提取物进行蛋白印迹分析。正如预期的一样,Histone H3 (K9M Mutant Specific) Antibody 仅检测 K9M 突变型组蛋白 H3.3,无法检测野生型 H3.3 蛋白。细胞系由马萨诸塞州总医院的 Konrad Hochedlinger 博士惠赠。
Western Blotting Image 2: Histone H3 (K9M Mutant Specific) Antibody Expand Image
使用 Histone H3(K9M 特定突变体)抗体(上图)、Myc-Tag (71D10) Rabbit mAb #2278(中间)和 Histone H3 (D1H2) XP® Rabbit mAb #4499(下图)对转染空载 (-) 或转染 (+) 带 Myc 标签的野生型组蛋白 H3 (Myc-Histone H3 WT) 或带 Myc 标签的 K9M 突变型组蛋白 H3 (Myc-Histone H3 K9M) 的 293T 细胞的提取物进行蛋白印迹分析。正如预计的一样,Histone H3(K9M 特定突变体)抗体仅检测带 Myc 标签的 K9M 突变型组蛋白 H3 蛋白,而不检测带 Myc 标签的野生型组蛋白 H3 或内源性野生型组蛋白 H3 蛋白。该抗体可检测 K9M 突变型组蛋白 H3 蛋白的内源水平。