| 货号 | 12799S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IHC-P/IF-IC/ChIP |
| 使用方法 | WB(1:1000) IP (1:100) IHC-P (1:1000) IF-IC (1:400) 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). The p300/CBP histone acetyltransferases acetylate multiple lysine residues in the amino terminal tail of histone H2B (Lys5, 12, 15, and 20) at gene promoters during transcriptional activation (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 access of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites that facilitate recruitment of many transcription and chromatin regulatory proteins that contain a bromodomain, which binds to acetylated lysine residues (6). Histone H2B is mono-ubiquitinated at Lys120 during transcriptional activation by the RAD6 E2 protein in conjunction with the BRE1A/BRE1B E3 ligase (also known as RNF20/RNF40) (7). Mono-ubiquitinated histone H2B Lys120 is associated with the transcribed region of active genes and stimulates transcriptional elongation by facilitating FACT-dependent chromatin remodeling (7-9). In addition, it is essential for subsequent methylation of histone H3 Lys4 and Lys79, two additional histone modifications that regulate transcriptional initiation and elongation (10). In response to metabolic stress, AMPK is recruited to responsive genes and phosphorylates histone H2B at Lys36, both at promoters and in transcribed regions of genes, and may regulate transcriptional elongation (11). In response to multiple apoptotic stimuli, histone H2B is phosphorylated at Ser14 by the Mst1 kinase (12). Upon induction of apoptosis, Mst1 is cleaved and activated by caspase-3, leading to global phosphorylation of histone H2B during chromatin condensation. Interestingly, histone H2B is rapidly phosphorylated at irradiation-induced DNA damage foci in mouse embryonic fibroblasts (13). In this case, phosphorylation at Ser14 is rapid, depends on prior phosphorylation of H2AX Ser139, and occurs in the absence of apoptosis, suggesting that Ser14 phosphorylation may have distinct roles in DNA-damage repair and apoptosis. |
| 存放说明 | -20C |
| 计算分子量 | 14 |
Confocal immunofluorescent analysis of HeLa cells, untreated (right) or treated with Trichostatin A (TSA) #9950 (left), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).采用共聚焦免疫荧光术检测untreated (左图)或treated with TSA #9950 (右图)的Hela细胞，使用的抗体为Acetyl-Histone H2B (Lys5) (D5H1S) XP®Rabbit mAb (绿色)。肌动蛋白微丝使用DY-554 鬼笔环肽进行标记（红色），假蓝色= DRAQ5® #4084 (DNA荧光染料)。 | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb in the presence of non-acetyl peptide (left) or Lys5 acetyl peptide (right).免疫组化检测石蜡包埋的人结肠癌，所使用的抗体为Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb，左图无乙酰化肽段存在，右图有Lys5位点乙酰化肽段存在。 | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb.免疫组化检测石蜡包埋的人乳腺癌，所使用的抗体为Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb。 | |
Western blot analysis of extracts from HeLa and C6 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18hr; +), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (upper) or Histone H2B (D2H6) Rabbit mAb #12364 (lower).Western blot检测untreated (-)或 treated with Trichostatin A (TSA) #9950 (1 μM, 18hr; +)的HeLa和C6细胞，使用的抗体为Acetyl-Histone H2B (Lys5) (D5H1S) XP®Rabbit mAb (上图) or Histone H2B (D2H6) Rabbit mAb #12364 (下图)。 | |
Immunoprecipitation of acetylated histone H2B from HeLa cell extracts treated with Trichostatin A (TSA) #9950 (1 μM, 18hr) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb.从 经Trichostatin A (TSA) #9950 (1 μM, 18hr) 处理的HeLa细胞提取物中免疫沉淀乙酰化的histone H2B，使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) 或 Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb (lane 3)。Lane 1为10% input。 使用Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb进行Western blot检测。 | |
Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated acetyl-histone H2B (Lys5) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H2B (Lys5) peptide competed away binding of the antibody.Peptide ELISA方法测定Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb 抗体特异性。 图片显示在不断提高不同竞争性多肽的浓度时抗体都能够与预包被好的acetyl-histone H2B (Lys5) peptide结合。同时也显示了，只有acetyl-histone H2B (Lys5) peptide能够竞争性抑制其与抗体的结合。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 µl of Acetyl-Histone H2B (Lys5) (D5H1S) 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 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 对4 x 106 HeLa细胞中交联过的染色质进行染色质免疫沉淀，所用抗体为10 µl of Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb 或2 µl Normal Rabbit IgG #2729，富集到的DNA经过实时PCR定量，所使用产品为SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP®Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, 以及SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。 |
