Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Methyl-Histone H3 Antibody Sampler Kit #9847

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725 40 µl W IP IHC-P IF-IC ChIP H M R Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys9) (D85B4) XP® Rabbit mAb #4658 40 µl W IP IF-IC ChIP H M R Mk (Dm) (X) (Z) (B) (Pg) (Sc) (Ce) 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys27) Antibody #9755 40 µl W IP H M R Mk 17 Rabbit
Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb #2901 40 µl W IHC-P IF-IC H M R Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys79) Antibody #9757 40 µl W H M R Mk 17 Rabbit
Histone H3 (D1H2) XP® Rabbit mAb #4499 40 µl W IHC-P IF-IC H M R Mk (Hm) (C) (Dm) (X) (Z) (B) 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  B=Bovine  Pg=Pig  Sc=S. cerevisiae  Ce=C. elegans
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

All antibodies in the Methyl-Histone H3 Antibody Sampler Kit recognize histone H3 only when modified at the indicated site.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys9) (D85B4) XP® Rabbit mAb #4658.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys27) Antibody #9755.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl Histone H3 (Lys79) Antibody #9757.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb #4499.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb #2901.


Description

The Methyl-Histone H3 Antibody Sampler Kit provides a fast and economical means of evaluating methylation sites on histone H3. The kit contains enough primary and secondary antibodies to perform four western blots.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic di-methylated peptides corresponding to residues surrounding Lys 4, 27 and 79 of human Histone H3. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic di-methylated peptides corresponding to residues surrounding Lys 9 and 36 of human Histone H3. Histone H3 (D1H2) XP Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein.

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.

Application References

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Protocols

* Product-specific protocol.

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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