| 货号 | 8647S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IHC-P/IF-IC/ChIP |
| 目标/特异性 | Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb recognizes endogenous levels of histone H4 protein only when acetylated at Lys5. This antibody does not cross-react with histone H4 acetylated at Lys8, Lys12, or Lys16. |
| 使用方法 | WB(1:1000) IP (1:100) IHC-P (1:6400) IF-IC (1:100) ChIP (1:25) |
| 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, 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 |
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (right) using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). 使用Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb (绿色)，共聚焦免疫荧光分析untreated (左图)或treated Trichostatin A (TSA) #9950 (右图)的HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。蓝色= DRAQ5® #4084 (DNA荧光染料)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 µl of Acetyl-Histone H4 (Lys5) (D12B3) 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, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, 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. 使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，来自4 x 106 HeLa细胞的交联染色质以及20 µl Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human MyoD1 Exon 1 Primers #4490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1的总input chromatin的数量的信号。 | |
Western blot analysis of extracts from various cell lines using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb. 使用Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb兔单抗，免疫印迹(Western blot)分析不同细胞中Acetyl-Histone H4 (Lys5) 蛋白水平。 | |
Western blot analysis of extracts from HeLa and C2C12 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18 hr; +), using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb (upper) or Histone H4 (L64C1) Mouse mAb #2935 (lower). 使用Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb (上图)或Histone H4 (L64C1) Mouse mAb #2935 (下图)，免疫印迹(Western blot)分析不同细胞中Acetyl-Histone H4 (Lys5)和Histone H4蛋白水平，细胞分为未处理组 (-)或处理组： Trichostatin A (TSA) #9950 (1 μM, 18 hr; +)。 | |
Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated acetyl-histone H4 (Lys5) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H4 (Lys5) peptide competed away binding of the antibody. 通过peptide ELISA确定Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb的特异性。该图描述了抗体与提前包被的acetyl-histone H4 (Lys5) 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acetyl-histone H4 (Lys5) peptide竞争脱离抗体的结合。 | |
Immunohistochemical analysis of paraffin-embedded human breast using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb in the presence of non-acetyl-histone H4 (Lys5) peptide (left) or acetyl-histone H4 (Lys5) peptide (right). 使用Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb，该抗体与non-acetyl-histone H4 (Lys5) peptide (左图)或acetyl-histone H4 (Lys5) peptide (右图)一起孵育，免疫组化分析人源乳腺癌组织石蜡切片。 | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb. 使用Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb兔单抗，免疫组化分析人源结肠癌组织石蜡切片。 |
