| 货号 | 5737S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/ChIP |
| 目标/特异性 | Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit mAb recognizes endogenous levels of histone H4 protein only when tri-methylated at Lys20. This antibody does not cross-react with non-methylated, mono-methylated or di-methylated histone H4 Lys20. This antibody detects a 95 kDa non-specific protein of unkown origin. |
| 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 | 11 |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Tri-Methyl-Histone H4 (Lys20) (D84D2) 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 α Satellite Repeat Primers #4486, SimpleChIP® Human AFM Intron 1 Primers #5098, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. 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 H4 (Lys20) (D84D2) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human α Satellite Repeat Primers #4486、SimpleChIP® Human AFM Intron 1 Primers #5098和SimpleChIP® Human GAPDH Exon 1 Primers #5516，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，这相当于一。 | |
Western blot analysis of extracts from HeLa and NIH/3T3 cells using Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit mAb. 使用Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit mAb兔单抗，免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Tri-Methyl-Histone H4 (Lys20)的蛋白水平。 | |
Tri-Methyl Histone H4 (Lys20) (D84D2) Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated tri-methyl histone H4 (Lys20) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the tri-methyl histone H4 (Lys20) peptide competed away binding of the antibody. 通过peptide ELISA确定Tri-Methyl Histone H4 (Lys20) (D84D2) Rabbit mAb兔单抗的特异性。该图描述了抗体与提前包被的tri-methyl histone H4 (Lys20) 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tri-methyl histone H4 (Lys20) peptide竞争脱离抗体的结合。 |
