Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb #2901

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IF-IC H M R Mk Endogenous 17 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb detects endogenous levels of histone H3 only when di-methylated on Lys36. The antibody does not cross-react with non-methylated, mono-methylated, or tri-methylated Lys36. In addition, the antibody does not cross-react with di-methylated histone H3 Lys4, Lys9, Lys27, Lys79 or methylated histone H4 Lys20.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys36 is di-methylated.

Western Blotting

Western Blotting

Antibody specificity was determined by Western blotting. HeLa and NIH/3T3 cell extracts were probed with Di-Methyl Histone H3 (Lys36) (C75H12) Rabbit mAb alone (Panel A) or Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb pre-adsorbed with 1.5 μM of various competitor peptides (Panels B-I). As shown, only the di-methyl-histone H3 (Lys36) peptide competed away binding of the antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).


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). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).

  1. Peterson, C.L. and Laniel, M.A. (2004) Curr. Biol. 14, R546-R551.
  2. Kubicek, S. et al. (2006) Ernst Schering Res. Found Workshop, 1-27.
  3. Lin, W. and Dent, S.Y. (2006) Curr. Opin. Genet. Dev. 16, 137-142.
  4. Lee, D.Y. et al. (2005) Endocr. Rev. 26, 147-170.
  5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-926.
  6. Shi, X. et al. (2006) Nature 442, 96-99.
  7. Wysocka, J. et al. (2006) Nature 442, 86-90.
  8. Wysocka, J. et al. (2005) Cell 121, 859-872.
  9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-217.

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