| 货号 | 27683S |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human\Mouse\Rat\Moneky |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IP , F , IF-IC , ChIP , ChIP-seq |
| 目标/特异性 | Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb recognizes endogenous levels of histone H3 protein only when acetylated at Lys36. This antibody does not cross react with other known acetylated lysine residues on histones H3, H4, H2A and H2B. |
| 使用方法 | W IP IF-IC F ChIP |
| 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 (1,2). Histone acetylation occurs mainly on the amino-terminal tail domains of histones H2A (Lys5), H2B (Lys5, 12, 15, and 20), H3 (Lys9, 14, 18, 23, 27, 36 and 56), and H4 (Lys5, 8, 12, and 16) and is important for the regulation of histone deposition, transcriptional activation, DNA replication, recombination, and DNA repair (1-3). Hyper-acetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the accessibility of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites for a protein module called the bromodomain, which binds to acetylated lysine residues (6). Many transcription and chromatin regulatory proteins contain bromodomains and may be recruited to gene promoters, in part, through binding of acetylated histone tails. Histone acetylation is mediated by histone acetyltransferases (HATs), such as CBP/p300, GCN5L2, PCAF, and Tip60, which are recruited to genes by DNA-bound protein factors to facilitate transcriptional activation (3). Deacetylation, which is mediated by histone deacetylases (HDAC and sirtuin proteins), reverses the effects of acetylation and generally facilitates transcriptional repression (7,8). |
| 存放说明 | -20C |
| 计算分子量 | 17 |
| 参考文献 | 1 . Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51. 2 . Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9. 3 . Roth, S.Y. et al. (2001) Annu Rev Biochem 70, 81-120. 4 . Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79. 5 . Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41. 6 . Yang, X.J. (2004) Bioessays 26, 1076-87. 7 . Haberland, M. et al. (2009) Nat Rev Genet 10, 32-42. 8 . Haigis, M.C. and Sinclair, D.A. (2010) Annu Rev Pathol 5, 253-95. 9 . Morris, S.A. et al. (2007) J Biol Chem 282, 7632-40. |
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (1 μM, 4 hrs; right) using Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). | |
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (green) using Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K-562 cells and either 10 μl of Acetyl-Histone H3 (Lys36) (D9T5Q) 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 β-Actin Promoter Primers #13653, SimpleChIP®Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human α Satellite Repeat Primers #4486, and SimpleChIP®Human MYT-1 Exon 1 Primers #4493. 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. | |
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18 hr; +), using Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K-562 cells and 10 μl of Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 50 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across RPL30, a known target gene of H3K36Ac (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet. |
