| 货号 | 9783T |
| 描述 | Tri-Methyl Histone H3 Antibody Sampler Kit offers an economical means to evaluate the tri-methylation of Histone H3 on multiple residues. The kit contains enough primary antibody to perform four western blot experiments per primary.Tri-Methyl Histone H3 Antibody Sampler Kit提供了一个快速和经济方法去评估histone H3蛋白三甲基化位点。该试剂盒包含充足的一抗和二抗去进行四次免疫印迹反应。 |
| 目标/特异性 | Each modification-state Histone H3 antibody detects endogenous levels of Histone H3 only when tri-methylated on the indicated lysine residue. These antibodies do not cross-react with mono-methylated or di-methylated histone H3, or tri-methylated histone H4 at Lys20. Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb detects endogenous levels of histone H3 when tri-methylated on Lys9. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys9, but does not cross-react with non-methylated or mono-methylated histone H3 Lys9. This antibody does not detect tri-methyl histone H3 Lys9 when the adjacent Ser10 residue is phosphorylated during mitosis. In addition, this antibody does not cross-react with methylated histone H3 Lys4, Lys27, Lys36, or Lys79. Tri-Methyl-Histone H3 (Lys79) Antibody may show slight cross-reactivity toward histone H3 when di-methylated at Lys79, but does not cross-react with histone H3 tri-methylated at Lys4, 9, 27, or 36, or histone H4 at Lys20. Histone H3 (D1H2) XP® Rabbit mAb detects endogenous levels of total histone H3 protein. This antibody does not cross-react with other histones. |
| 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).核小体是由四种组蛋白(H2A、H2B、H3和H4)组成，它是染色质的主要构成模块。起初被认为作为一个DNA包装的静态支架，现在则显示组蛋白是动态蛋白，经历多种翻译后修饰的形式，包括乙酰化、磷酸化、甲基化和泛素化(1)。组蛋白甲基化对于该基因组的活化和未活化区域的形成有着主要决定作用，并且在发育期间对该基因组的正确规划起着关键作用(2,3)。histones H3 (Arg2、17、26)和H4 (Arg3)的精氨酸甲基化促进转录调控以及通过蛋白质精氨酸甲基转移酶(PRMTs)家族的介导，包括共激活因子PRMT1和CARM1 (PRMT4) (4)。相反，多种多样的组蛋白赖氨酸甲基转移酶已经被鉴定，除了这个之外其它的都包含一个保守的催化SET区域，这个起初被鉴定在Drosophila Su(var)3-9、zeste增强子和Trithorax蛋白。赖氨酸甲基化主要发生在histones H3 (Lys4、9、27、36、79)和H4 (Lys20)，并且已经涉及到转录激活和沉默(4)。这些赖氨酸残基的甲基化协调染色质修饰酶的招募包括methyl-lysine结合模块例如chromodomains (HP1, PRC1)、PHD fingers (BPTF, ING2)、tudor domains (53BP1)和WD-40 domains (WDR5) (5-8)。组蛋白例如PADI4、LSD1、JMJD1、JMJD2和JHDM1的发现已经显示甲基化是一个可逆的后生标记物(9)。 |
| 存放说明 | -20C |
| 参考文献 | Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42. Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26. Shi, X. et al. (2006) Nature 442, 96-9. Wysocka, J. et al. (2006) Nature 442, 86-90. Wysocka, J. et al. (2005) Cell 121, 859-72. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7. |
Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb (green) and COX IV (4D11-B3-E8) Mouse mAb #11967 (red). | |
Antibody specificity was determined by western blotting. HeLa and NIH/3T3 cell lysates were probed with Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb (Panel A) or Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb pre-adsorbed with 1.5 μM of various competitor peptides (panels B-M). As shown, only the tri-methyl histone H3 (Lys9) peptide (panel E) competed away binding of the antibody. | |
Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb. | |
Confocal immunofluorescent analysis of interphase (left) or mitotic (right) HeLa cells, untreated (upper) or λ phosphatase-treated (lower), using Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). As shown, this antibody does not detect tri-methyl histone H3 Lys9 in mitotic cells when the adjacent Ser10 residue is phosphorylated. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Tri-Methyl-Histone H3 (Lys9) (D4W1U) 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 Exon 1 Primers #5516, SimpleChIP® Human AFM Intron 1 Primers #5098, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, 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. | |
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. | |
Flow cytometric analysis of human whole blood cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) 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 lymphoma using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb in the presence of non-methyl peptide (left) or K27 tri-methyl peptide (right). | |
Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to 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. | |
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). | |
Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb #4499. | |
Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys79) Antibody #4260. | |
Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb #4909. | |
Western blot analysis of extracts from various cell lines using Tri-Methyl Histone H3 (Lys27) (C36B11) Rabbit mAb #9733. | |
Western blot analysis of extracts from various cell lines using Tri-Methyl Histone H3 (Lys9) Anibody #9754. |
