Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647 Conjugate) #12158

conjugate   h3   H3K27   H3K27me3   histone  

No. Size Price
12158S 100 µl ( 50 tests ) ¥3,986.00 现货查询 购买询价
12158 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse,Rat,Monkey, Endogenous Rabbit IgG
IF-IC 1:50

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Xenopus, Zebrafish,

Specificity / Sensitivity

Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of histone H3 only when tri-methylated at Lys27. The antibody does not cross-react with non-methylated, mono-methylated or di-methylated Lys27. In addition, the antibody does not cross-react with mono-methylated, di-methylated or tri-methylated histone H3 at Lys4, Lys9, Lys36 or histone H4 at Lys20.

Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb 兔单抗(Alexa Fluor® 647 Conjugate)能够检测内源性的Lys27位点三甲基化的组蛋白H3。该抗体不会与Lys27位点非甲基化,单甲基化或双甲基化组蛋白H3发生交叉反应。此外,也不会与Lys4,Lys9,Lys36 位点单甲基化,二甲基化或三甲基化的组蛋白H3或Lys20位点单、二或三甲基化的组蛋白 H4发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is tri-methylated.

该单克隆抗体是经由合成的Lys27位点三甲基化组蛋白H3的氨基端肽段免疫动物而生产的。

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733.

该CST抗体偶联有Alexa Fluor®647荧光染料,通过直标法流式细胞术和免疫荧光分析在人细胞内进行内部检测。它与未偶联的Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733 具有相同的种属交叉反应性。

Flow Cytometry

Flow Cytometry

Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Alternate Protocol and stained using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647 Conjugate). The forward/side-scatter lymphocyte gate was applied to a histogram depicting the mean fluorescence intensity of Tri-Methyl-Histone H3 (Lys27) (blue) and concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (red).按照CST 流式替代操作方法对人全血进行固定,溶解以及透性化处理,使用Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647 Conjugate)进行染色。采用前向角/侧向角散射光设置淋巴细胞门,以直方图来表示Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor®647偶联)(蓝色)和浓度匹配的Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (红色) 的平均荧光强度。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor). Actin filaments were labeled with DY-554 phalloidin (red).采用共聚焦免疫荧光术检测Hela细胞,使用的抗体为Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 647偶联) (蓝色 假彩色)。肌动蛋白微丝使用DY-554 鬼笔环肽进行标记(红色)。

Background

The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).

核小体是构成染色质的基本结构单位,由四种核心组蛋白(H2A、H2B、H3和H4)组成。最初认为组蛋白的功能是作为DNA包装的静态骨架,但现在研究表明,组蛋白是一个动态的蛋白,参与了多种类型的翻译后修饰,包括乙酰化,磷酸化,甲基化以及泛素化(1)。组蛋白甲基化是基因组活动和非活动区域形成的一个重要决定因素,同时对发育过程中基因组的正确编程也是至关重要的(2,3)。位于组蛋白H3(Arg2,17,26)和H4 (Arg3)的精氨酸甲基化能够促进转录激活,该甲基化作用是由蛋白精氨酸甲基转移酶 (PRMTs)家族催化的,其中包括共激活因子 PRMT1 和 CARM1 (PRMT4)(4)。与之不同的是,组蛋白赖氨酸甲基转移酶家族更多样化,除了其中一个,其它所有成员都包含一个保守的SET催化结构域,该SET域(Su(var)3-9, Enhancer of zeste以及Trithorax蛋白)最初是在果蝇中得到确认的。赖氨酸甲基化主要发生于组蛋白 H3 (Lys4,9、 27、 36、 79) 和 H4 (Lys20), 与转录激活和沉默都直接相关 (4)。这些赖氨酸残基的甲基化能够协调染色质修饰酶的募集,这些染色质修饰酶都含有甲基化赖氨酸结合模块,比如染色质结构域(HP1, PRC1), PHD指纹 (BPTF, ING2), tudor 结构域 (53BP1)以及 WD-40 结构域(WDR5) (5-8)。经过研究,人们发现了如PADI4,LSD1,JMJD1,JMJD2 以及 JHDM1等组蛋白去甲基化酶,这些都表明甲基化修饰是一个可逆的表观遗传学标志。

  1. Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.
  2. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
  3. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
  4. Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.
  5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.
  6. Shi, X. et al. (2006) Nature 442, 96-9.
  7. Wysocka, J. et al. (2006) Nature 442, 86-90.
  8. Wysocka, J. et al. (2005) Cell 121, 859-72.
  9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.

Application References

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Protocols

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

U.S. Patent No. 5,675,063.

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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