Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb #12522

H3   H3K79me1   H3Lys79   Histone  

No. Size Price
12522S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
12522 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 17 Rabbit IgG
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, ChIP=Chromatin IP,

Specificity / Sensitivity

Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb recognizes endogenous levels of histone H3 protein only when mono-methylated at Lys79. This antibody does not cross-react with non-methylated, di-methylated, or tri-methylated histone H3 Lys79. In addition, the antibody does not cross-react with histone H3 mono-methylated at Lys4, Lys9, Lys27, or Lys36.

Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb兔单抗能够检测内源性的Lys79位点单甲基化的组蛋白H3。该抗体不会与Lys79位点非甲基化,二或三甲基化的组蛋白H3发生交叉反应。此外,也不会与Lys4、Lys9、Lys27或Lys36 位点单甲基化的组蛋白H3发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding mono-methyl Lys79 of human histone H3 protein.

该单克隆抗体采用合成的与人源Lys79位点单甲基化组蛋白H3的氨基端肽段免疫动物而生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb.mAb.

Western blot方法检测不同细胞系的提取物,所用抗体为Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb兔单抗。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 µl of Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Promoter Primers #4471, SimpleChIP® Human GAPDH Intron 2 Primers #4478, human PABPC1 promoter primers, and human PABPC1 intron 4 primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

使用4 x 106 HeLa细胞中交联过的染色质以及10 µl Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀,所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量,所使用产品为SimpleChIP® Human GAPDH Promoter Primers #4471、SimpleChIP® Human GAPDH Intron 2 Primers #4478、human PABPC1 promoter primers以及 human PABPC1 intron 4 primers。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。

ELISA-Peptide

ELISA-Peptide

Mono-Methyl Histone H3 (Lys79) (D5X1S) Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated mono-methyl histone H3 (Lys79) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the mono-methyl histone H3 (Lys79) peptide competed away binding of the antibody.

Peptide ELISA方法测定Mono-Methyl Histone H3 (Lys79) (D5X1S) Rabbit mAb兔单抗 抗体特异性。 图片显示在不断提高不同竞争性多肽的浓度时抗体都能够与预包被好的mono-methyl histone H3 (Lys79) peptide结合。同时也可以看出,只有mono-methyl histone H3 (Lys79) peptide能够竞争性抑制其与抗体的结合。

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.

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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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