| 货号 | 9751T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human,Mouse,Rat,Monkey,D. melanogaster,S. cerevisiae, |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IHC-P , F , 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) IHC-P (1:200) F (1:200) IF-IC (1:400) ChIP (1:50) |
| 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. |
Flow cytometric analysis of human whole blood cells using Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (blue) and Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. Analysis was performed on cells in the lymphocyte gate. | |
Immunohistochemical analysis of paraffin-embedded human colon using Tri-Methyl-Histone H3 (K4) (C42D8) Rabbit mAb in the presence of non-methyl peptide (left) or K4 tri-methyl peptide (right). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Tri-Methyl-Histone H3 (Lys4) (C42D8) 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 RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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10 µl Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human GAPDH Exon 1 Primers #5516、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。 | |
Antibody specificity was determined by Western blotting. HeLa and NIH/3T3 cell lysates were probed with Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb (Panel A) or Tri-Methyl Histone H3 (Lys4) Rabbit mAb pre-adsorbed with 1.5 μM of various competitor peptides (Panels B-M). As shown, only the tri-methyl histone H3 (Lys4) peptide competed away binding of the antibody. 抗体特异性通过免疫印迹(Western blot)确定。HeLa和NIH/3T3细胞提取物的探针使用只有Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb (图A)或Tri-Methyl Histone H3 (Lys4) Rabbit mAb，并且其提前孵育了1.5 μM various competitor peptides (图B-I)。如图所示，仅tri-methyl histone H3 (Lys4) peptide竞争离开该抗体的结合。 | |
Western blot analysis of various cell types using Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb. 使用Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb，免疫印迹(Western blot)分析不同细胞中Tri-Methyl-Histone H3 (Lys4)的蛋白水平。 | |
Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). 使用Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (绿色)，共聚焦免疫荧光分析HeLa细胞。Alexa Fluor® 555 phalloidin标记微丝蛋白(红色)。 |
