Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb #5327

H3   H3.1   H3K9   H3K9me2   H3k9me3   histone  

No. Size Price
5327S 100 µl ( 20 western blots ) ¥3,900.00 现货查询 购买询价
5327T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
5327 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:2000 Human,Mouse,Rat,Monkey, Endogenous 17 Mouse IgG1
IP 1:100
IF-IC 1:100
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb detects endogenous levels of histone H3 when di- or tri-methylated on Lys9. The antibody also shows slight cross-reactivity with histone H3 when mono-methylated on Lys9. The antibody does not cross-react with methylated histone H3 Lys4, Lys27, Lys36 or Lys79.

Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb鼠单抗能够检测仅在Lys9位点双甲基化或三甲基化的内源性histone H3的蛋白水平。该抗体也显示与Lys9位点单甲基化histone H3发生弱交叉反应。该抗体不与Lys4、Lys27、Lys36或Lys79位点甲基化的histone H3蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is tri-methylated.

通过人工合成仅在lys9位点三甲基化histone H3蛋白氨基端相应的多肽片段去免疫动物从而制备出单克隆抗体。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb or 2 μl 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 Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human α Satellite Repeat Primers #4486, and SimpleChIP® Human AFM Intron 1 Primers #5098. 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SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,用4 x 106 HeLa细胞的交联染色质以及10 µl Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human α Satellite Repeat Primers #4486和SimpleChIP® Human AFM Intron 1 Primers #5098,浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号,相当于一个。



Confocal immunofluorescent analysis of HeLa cells using Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

使用Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb 鼠单抗(绿色),共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cell lines using Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb.

使用Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb鼠单抗,免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Di/Tri-Methyl-Histone H3 (Lys9)的蛋白水平。



Di/Tri-Methyl Histone H3 (Lys9) (6F12) Mouse mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated tri-methyl histone H3 (Lys9) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl and tri-methyl histone H3 (Lys9) peptides compete away binding of the antibody.

通过peptide ELISA确定Di/Tri-Methyl Histone H3 (Lys9) (6F12) Mouse mAb鼠单抗的特异性。该图描述了抗体与提前包被的tri-methyl histone H3 (Lys9) peptide的结合能力,并且多肽中含有浓度递增的不同竞争多肽。如同所示,仅di-methyl and tri-methyl histone H3 (Lys9) peptides竞争脱离抗体的结合。


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).

核小体是由四种组蛋白(H2A、H2B、H3和H4)组成,它是染色质的主要构成模块。起初被认为作为一个DNA包装的静态支架,现在则显示组蛋白是动态蛋白,经历多种翻译后修饰的形式,包括乙酰化、磷酸化、甲基化和泛素化(1)。组蛋白甲基化对于该基因组的活化和未活化区域的形成有着主要决定作用,并且在发育期间对该基因组的正确规划起着关键作用(2,3)。histones H3 (Arg2、17、26)和H4 (Arg3)的精氨酸甲基化促进转录调控以及通过蛋白质精氨酸甲基转移酶(PRMTs)家族的介导,包括共激活因子PRMT1和CARM1 (PRMT4) (4)。相反,多种多样的组蛋白赖氨酸甲基转移酶已经被鉴定,除了这个之外其它的都包含一个保守的催化SET区域,这个起初被鉴定在Drosophila Su(var)3-9、zeste增强子和Trithorax蛋白。赖氨酸甲基化主要发生在histones H3 (Lys4、9、27、36、79)和H4 (Lys20),并且已经涉及到转录激活和沉默(4)。这些赖氨酸残基的甲基化协调染色质修饰酶的招募包括methyl-lysine结合模块例如chromodomains (HP1, PRC1)、PHD fingers (BPTF, ING2)、tudor domains (53BP1)和WD-40 domains (WDR5) (5-8)。组蛋白例如PADI4、LSD1、JMJD1、JMJD2和JHDM1的发现已经显示甲基化是一个可逆的表遗传标记物(9)。

  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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