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45139
Methyl-Histone H3 (Lys79) Antibody Sampler Kit
Primary Antibodies

Methyl-Histone H3 (Lys79) Antibody Sampler Kit #45139

Western Blotting Image 1

Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb.

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Chromatin IP-seq Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of H3K79me3 (see additional figures containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 3

Western blot analysis of extracts from HeLa and NIH/3T3 cell lines using Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb.

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Western Blotting Image 4

Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb.

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Western Blotting Image 5

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

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Western Blotting Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Chromatin IP Image 7

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Promoter Primers #4471, and SimpleChIP® Human MyoD1 Exon 1 Primers #4490. 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.

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Chromatin IP Image 8

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb or 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 γ-Actin Promoter Primers #5037, SimpleChIP® Human γ-Actin Intron 3 Primers #5047, SimpleChIP® Human GAPDH Promoter Primers #4471, and SimpleChIP® Human GAPDH Intron 2 Primers #4478. 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.

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Chromatin IP Image 9

Chromatin immunoprecipitations were performed with cross-linked chromatin fromHeLa cells and either Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb or 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 Promoter Primers #4471, SimpleChIP® Human GAPDH Intron 2 Primers #4478, human PABPC1 promoter primers, and human PABPC1 intron 4 primers. 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.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.

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Flow Cytometry Image 11

Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

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IF-IC Image 12

Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb 74073 20 µl
  • WB
  • ChIP
H M R Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb 5427 20 µl
  • WB
  • ChIP
H M R Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb 12522 20 µl
  • WB
  • ChIP
H M R Mk 17 Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Methyl-Histone H3 (Lys79) Antibody Sampler Kit provides an economical means of detecting levels of mono-, di-, and tri-methyl histone H3 Lys79 using methyl-specific and control histone H3 antibodies. The kit contains enough primary antibodies to perform at least two western blot experiments.

Each antibody in the Methyl-Histone H3 (Lys79) Antibody Sampler Kit detects endogenous levels of its target protein. Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb recognizes endogenous levels of histone H3 protein when tri-methylated at Lys79. This antibody shows some cross-reactivity to histone H3 that is di-methylated on Lys79, but does not cross-react with non-methylated or mono-methylated histone H3 Lys79. Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb detects endogenous levels of histone H3 only when di-methylated on Lys79. Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb recognizes endogenous levels of histone H3 protein only when mono-methylated at Lys79. These methyl-Lys79 antibodies do not cross-react with histone H3 methylated at Lys4, Lys9, Lys27, or Lys36. Histone H3 (D1H2) XP® Rabbit mAb detects endogenous levels of total Histone H3 protein, including isoforms H3.1, H3.2, H3.3, and the variant histone CENP-A. This antibody does not cross-react with other core histones.

In addition, the antibody does not cross-react with methylated histone H3 Lys4, Lys9, Lys27, Lys36 or methylated histone H4 Lys20.

Monoclonal methyl-histone H3 Lys79 antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of histone H3 in which Lys79 is mono-, di-, or tri-methylated. The control histone H3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein.

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).

Methylation of histone H3 Lys79 is mediated by the DOT1L histone methyltransferase, is associated with transcriptionally active genes, and plays a role in the regulation of DNA damage response, cell cycle, and embryonic stem cell development.

  1. Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.
  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, 137-42.
  4. Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.
  5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.
  6. Shi, X. et al. (2006) Nature 442, 96-9.
  7. Wysocka, J. et al. (2006) Nature 442, 86-90.
  8. Wysocka, J. et al. (2005) Cell 121, 859-72.
  9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.
Entrez-Gene Id
8350
Swiss-Prot Acc.
P68431
For Research Use Only. Not For Use In Diagnostic Procedures.

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XP is a registered trademark of Cell Signaling Technology, Inc.

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