| 货号 | 8240T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human,Mouse,Rat,Monkey, |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IP , F , IF-IC , ChIP |
| 目标/特异性 | Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb recognizes endogenous levels of histone H2A protein only when ubiquitinated at Lys119. The antibody does not cross-react with other ubiquitinated proteins or free ubiquitin. |
| 使用方法 | WB(1:2000) IP (1:100) F (1:1600) IF-IC (1:1600) 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). Ubiquitin is a conserved 76 amino acid peptide unit that can be covalently linked to many cellular proteins by the ubiquitination process. Three components are involved in this protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thioester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (2). Histone H2A is mono-ubiquitinated at Lys119 by the Polycomb Repressor Complex 1 (PRC1) and is critical for transcriptional silencing of the developmental HOX genes and X chromosome inactivation (3-6). PRC1 is composed of Bmi1 and RING1A (also RING1 or RNF1), both of which act to enhance the E3 ubiquitin ligase activity of the catalytic subunit RING1B (also RING2 or RNF2) (3,4). Histone H2A is also mono-ubiquitinated at Lys119 at sites of DNA damage. This mono-ubiquitination event requires the PRC1 components Bmi1 and RING1B, in addition to another E3 ubiquitin ligase RNF8, and contributes to subsequent recruitment of the BRCA1 complex, via binding of RAP80/UIMC1 (ubiquitin interactive motif containing 1 protein) (7-10). |
| 存放说明 | -20C |
| 计算分子量 | 23 |
| 参考文献 | 1 . Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51. 2 . Liu, F. and Walters, K.J. (2010) Trends Biochem Sci 35, 352-60. 3 . Wang, H. et al. (2004) Nature 431, 873-8. 4 . Cao, R. et al. (2005) Mol Cell 20, 845-54. 5 . de Napoles, M. et al. (2004) Dev Cell 7, 663-76. 6 . Fang, J. et al. (2004) J Biol Chem 279, 52812-5. 7 . Ginjala, V. et al. (2011) Mol Cell Biol 31, 1972-82. 8 . Bergink, S. et al. (2006) Genes Dev 20, 1343-52. 9 . Marteijn, J.A. et al. (2009) J Cell Biol 186, 835-47. 10 . Wu, J. et al. (2009) Mol Cell Biol 29, 849-60. |
Confocal immunofluorescent analysis of HeLa cells using Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb (green) and COX IV (4D11-B3-E8) Mouse mAb #11967 (red). | |
Western blot analysis of extracts from various cell lines using Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb. 使用Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb兔单抗，免疫印迹(Western blot)分析不同细胞系中Histone H2A的蛋白水平。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 µl of Ubiquityl-Histone H2A (Lys119) (D27C4) 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 human HoxA1 intron 1 primers, SimpleChIP® Human HoxA2 Promoter Primers #5517, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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 (equivalent to one). 使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，来自4 x 106 NCCIT细胞的交联染色质以及10 µl Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用human HoxA1 intron 1 primers、SimpleChIP® Human HoxA2 Promoter Primers #5517、SimpleChIP® Human GAPDH Exon 1 Primers #5516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（这相当于一）。 |
