Product Pathways - Chromatin Regulation / Epigenetics
Acetyl-Histone H2B (Lys12) (D7H4) Rabbit mAb (ChIP Formulated) #9072
|9072S||100 µl (10 immunoprecipitations)||---||In Stock||---|
|9072||carrier free and custom formulation / quantity||email request|
Species cross-reactivity is determined by western blot.
Applications Key: IP=Immunoprecipitation, ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Mouse, Rat, Monkey, Chicken, Xenopus, Zebrafish, Bovine, Pig, Horse.
Specificity / Sensitivity
Acetyl-Histone H2B (Lys12) (D7H4) Rabbit mAb (ChIP Formulated) recognizes endogenous levels of histone H2B protein only when acetylated at Lys12. This antibody does not cross-react with histone H2B acetylated at Lys5, Lys15, or Lys20.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic acetylated peptide corresponding to residues surrounding Lys12 of human histone H2B protein.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 µl of Acetyl-Histone H2B (Lys12) (D7H4) Rabbit mAb (ChIP Formulated) 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.
Acetyl-Histone H2B (Lys12) (D7H4) Rabbit mAb (ChIP Formulated) specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated acetyl-histone H2B (Lys12) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H2B (Lys12) peptide competed away binding of the antibody.
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.
- Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.
- Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
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- Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
- Yang, X.J. (2004) Bioessays 26, 1076-87.
- Kim, J. et al. (2009) Cell 137, 459-71.
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- Shilatifard, A. (2006) Annu Rev Biochem 75, 243-69.
- Bungard, D. et al. (2010) Science 329, 1201-5.
- Cheung, W.L. et al. (2003) Cell 113, 507-17.
- Fernandez-Capetillo, O. et al. (2004) J Exp Med 199, 1671-7.
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
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