| 货号 | 12522S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/ChIP |
| 目标/特异性 | 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. |
| 使用方法 | WB(1:1000) ChIP (1:50) |
| 供应商 | CST |
| 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 |
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 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。 | |
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。 |
