| 货号 | 5326S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IF-IC/ChIP |
| 目标/特异性 | Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb detects endogenous levels of histone H3 only when mono-methylated on Lys4. The antibody does not cross-react with non-methylated, di-methylated or tri-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) IF-IC (1:800) 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 |
Confocal immunofluorescent analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb #4694 (blue). Actin filaments were labeled with Dy-554 phalloidin (red). 使用Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb 兔单抗(绿色)和MEK1/2 (L38C12) Mouse mAb #4694 鼠单抗(蓝色)，共聚焦免疫荧光分析HeLa细胞。Dy-554 phalloidin标记微丝蛋白(红色)。 | |
Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb. 使用Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb兔单抗，免疫印迹(Western blot)分析不同细胞中Mono-Methyl-Histone H3 (Lys4)的蛋白水平。 | |
Mono-Methyl Histone H3 (Lys4) (D1A9) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated mono-methyl histone H3 (Lys4) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the mono-methyl histone H3 (Lys4) peptide competed away binding of the antibody. 通过peptide ELISA确定Mono-Methyl Histone H3 (Lys4) (D1A9) XP® Rabbit mAb兔单抗的特异性。该图描述了抗体与提前包被的mono-methyl histone H3 (Lys4) 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mono-methyl histone H3 (Lys4) peptide竞争脱离抗体的结合。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K562 cells and either 10 μl of Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® 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 ADAM9 Intron 11 Primers #73401, human ADAM18 intron 14 primers, human Trio inton 1 primers, and SimpleChIP®Human Trio Exon 57 Primers #90568. 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. |
