Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone H3 (Lys36) Antibody #11885

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Mk (Hm) (X) (Z) (GP) (Hr) Endogenous 17 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  X=Xenopus  Z=Zebrafish  GP=Guinea Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Acetyl-Histone H3 (Lys36) Antibody recognizes endogenous levels of histone H3 protein only when acetylated at Lys36. This antibody does not cross react with histone H3 acetylated at Lys9, 14, 18, 23, 27, or 56.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding acetylated Lys36 of human histone H3 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

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 H3 (Lys36) Antibody (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

ELISA-Peptide

ELISA-Peptide

Acetyl-Histone H3 (Lys36) Antibody specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to precoated acetyl-histone H3 (Lys36) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H3 (Lys36) peptide competed away binding of the antibody.

Background

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).

Histone H3 Lys36 acetylation is mediated by the GCN5 histone acetyltransferase and is conserved in multiple species, from yeast to mammals (9). Acetyl-histone H3 Lys36 is localized predominantly to the promoters of active RNA polymerase II-transcribed genes and overlaps with other acetylation marks associated with transcriptional activation, such as acetyl-histone H3 Lys9 and Lys14 (9). The pattern of acetyl-histone H3 Lys36 is inversely related to that of tri-methyl-histone H3 Lys36, which is found in the gene body of actively transcribed genes.

  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.

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