| 货号 | 9727L |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IHC-P/IF-IC/ChIP |
| 目标/特异性 | Tri-Methyl-Histone H3 (Lys4) Antibody detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, the antibody does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20. |
| 使用方法 | WB(1:1000) IP (1:50) IHC-P (1:200) IF-IC (1:4000) ChIP (1:100) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 17 |
| 参考文献 | 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. |
Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells, using Tri-Methyl-Histone H3 (Lys4) antibody. 使用Tri-Methyl-Histone H3 (Lys4) antibody，免疫印迹(Western blot)分析HeLa、NIH/3T3、C6和COS细胞中Tri-Methyl-Histone H3 (Lys4)的蛋白水平。 | |
Tri-Methyl-Histone H3 (Lys4) Antibody specificity was determined by peptide ELISA. Each graph depicts a titration of this antibody and the corresponding reactivity toward the non-methyl, mono-methyl, di-methyl and tri-methyl states of the indicated histone H3 or H4 lysine residue. 通过peptide ELISA确定Tri-Methyl-Histone H3 (Lys4) Antibody的特异性。每个图描述了该抗体的滴度和指定的histone H3或H4赖氨酸残基的非甲基化、单甲基化、双甲基化和三甲基化相应的活性。 | |
Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Tri-Methyl-Histone H3 (Lys4) Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). 与isotype control (右图)比较，使用Tri-Methyl-Histone H3 (Lys4) Antibody (绿色, 左图)，共聚焦免疫荧光分析NIH/3T3细胞。Alexa Fluor® 555 phalloidin标记微丝蛋白(红色)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Tri-Methyl-Histone H3 (Lys4) Antibody 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 Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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5 µl Tri-Methyl-Histone H3 (Lys4) Antibody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 MyoD1 Exon 1 Primers #4490和SimpleChIP® Human α Satellite Repeat Primers #4486，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。 | |
Immunohistochemical analysis of paraffin-embedded human colon using Tri-Methyl-Histone H3 (K4) Rabbit Antibody in the presence of non-methyl peptide (left) or K4 tri-methyl peptide (right). 使用Tri-Methyl-Histone H3 (K4) Rabbit Antibody免疫组化分析人结肠癌组织石蜡切片，左图： 非甲基化肽段，右图：K4 tri-methyl peptide。 |
