Tri-Methyl-Histone H3 (Lys4) AntibodyProduct information
Product Pathways - Chromatin Regulation / Epigenetics
Tri-Methyl-Histone H3 (Lys4) Antibody #9727
|9727L||300 µl (30 western blots)||---||In Stock||---|
|9727S||100 µl (10 western blots)||---||In Stock||---|
|9727||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Rat, Monkey||17||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Xenopus, Zebrafish.
Specificity / Sensitivity
Tri-Methyl-Histone H3 (Lys4) Antibody detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, the antibody does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which lysine 4 is tri-methylated. Antibodies are purified by protein A and peptide affinity chromatography.
Tri-Methyl-Histone H3 (Lys4) Antibody specificity was determined by peptide ELISA. Each graph depicts a titration of this antibody and the corresponding reactivity toward the non-methyl, mono-methyl, di-methyl and tri-methyl states of the indicated histone H3 or H4 lysine residue.
Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells, using Tri-Methyl-Histone H3 (Lys4) antibody.
Immunohistochemical analysis of paraffin-embedded human colon using Tri-Methyl-Histone H3 (K4) Rabbit Antibody in the presence of non-methyl peptide (left) or K4 tri-methyl peptide (right).
Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Tri-Methyl-Histone H3 (Lys4) Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Tri-Methyl-Histone H3 (Lys4) Antibody 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.
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).
- Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.
- Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
- Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
- Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.
- Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.
- Shi, X. et al. (2006) Nature 442, 96-9.
- Wysocka, J. et al. (2006) Nature 442, 86-90.
- Wysocka, J. et al. (2005) Cell 121, 859-72.
- Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.
- Song, N. et al. (2011) Acta Histochem Cytochem 44, 183-90. Applications: IHC-P (paraffin).
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 15019 Animal-Free Blocking Solution (5X)
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 9725 Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb
- 14166 Hematoxylin
- 9717 Histone H3 (3H1) Rabbit mAb
- 4499 Histone H3 (D1H2) XP® Rabbit mAb
- 4620 Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated)
- 9715 Histone H3 Antibody
- 2650 Histone H3 Antibody (ChIP Formulated)
- 13079 Immunohistochemistry Application Solutions Kit (Rabbit)
- 9707 Methyl-Histone H3 (Arg2) Antibody
- 9723 Mono-Methyl-Histone H3 (Lys4) Antibody
- 9701 Phospho-Histone H3 (Ser10) Antibody
- 6883 SignalFire™ ECL Reagent
- 12757 SignalFire™ Elite ECL Reagent
- 12630 SignalFire™ Plus ECL Reagent
- 14746 SignalStain® Citrate Unmasking Solution (10X)
- 14177 SignalStain® Mounting Medium
- 1064 Tri-Methyl-Histone H3 (Lys4) Blocking Peptide
- 9997 Tris Buffered Saline with Tween® 20 (TBST-10X)
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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.