| 货号 | 13944S |
| 反应种属 | Human/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 使用方法 | WB(1:1000) IP (1:50) F (1:1600) IF-IC (1:1600) 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 (1,2). 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 | 11 |
Western blot analysis of extracts from HeLa and C6 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 hr), using Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb (upper) and Histone H4 (D2X4V) Rabbit mAb #13919 (lower). | |
Confocal immunofluorescent analysis of HeLa cells untreated (left) or treated (right) with Trichostatin A (TSA) #9950 (1 µM, 4 hr) using Acetyl-Histone H4 (Lys12) (D2W60) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Acetyl-Histone H4 (Lys12) (D2W6O) 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 RPL30 Exon 3 Primers #7014, 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 HeLa cells, untreated (blue) or treated with SAHA (green) using Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab)2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as secondary antibody. |
