New England Biolabs Canada
 

Product Pathways - Chromatin Regulation / Epigenetics

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

Item# Description List Price Web Price Qty
45139T Methyl-Histone H3 (Lys79) Antibody Sampler Kit - 1 Kit $444.00
$399.60
ADD TO CART
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
X

Companion Products

Item# Description List Price Web Price Qty
12522S Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb - 100 µl $424.00
$381.60
ADD TO CART
4499L Histone H3 (D1H2) XP® Rabbit mAb - 300 µl $973.00
$875.70
ADD TO CART
4499S Histone H3 (D1H2) XP® Rabbit mAb - 100 µl $410.00
$369.00
ADD TO CART
4499T Histone H3 (D1H2) XP® Rabbit mAb - 20 µl $171.00
$153.90
ADD TO CART
5427S Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb - 100 µl $470.00
$423.00
ADD TO CART
5427T Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb - 20 µl $188.00
$169.20
ADD TO CART
7003P 20X LumiGLO® Reagent and 20X Peroxide - 5 ml each $90.00
$81.00
ADD TO CART
7003S 20X LumiGLO® Reagent and 20X Peroxide - 25 ml each $287.00
$258.30
ADD TO CART
7071S Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody - 50 assays $395.00
$355.50
ADD TO CART
7074P2 Anti-rabbit IgG, HRP-linked Antibody - 100 µl $106.00
$95.40
$79.50
ADD TO CART
7074S Anti-rabbit IgG, HRP-linked Antibody - 1 ml $199.00
$179.10
$149.25
ADD TO CART
7074V Anti-rabbit IgG, HRP-linked Antibody - 5 ml $633.00
$569.70
$474.75
ADD TO CART
74073S Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb - 100 µl $424.00
$381.60
ADD TO CART
7727L Biotinylated Protein Ladder Detection Pack - 3250 µl $319.00
$287.10
ADD TO CART
7727S Biotinylated Protein Ladder Detection Pack - 650 µl $90.00
$81.00
ADD TO CART
9998S BSA - 50 g $307.00
$276.30
ADD TO CART
9999S Nonfat Dry Milk - 250 g $84.00
$75.60
ADD TO CART
VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb #74073 20 µl W, ChIP, ChIP-seq H, M, R, Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb #5427 20 µl W, ChIP H, M, R, Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb #12522 20 µl W, ChIP, ChIP-seq H, M, R, Mk 17 Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb #4499 20 µl W, IHC-P, IF-IC, F H, M, R, Mk Hm, C, Dm, X, Z, B 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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


Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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

Chromatin IP-seq

Chromatin IP-seq

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.


Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Mono-Methyl-Histone H3 (Lys79) (D5X1S) 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 RAB5C gene. For additional ChIP-seq tracks, please download the product data sheet.

IHC-P (paraffin)

IHC-P (paraffin)

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

Chromatin IP

Chromatin IP

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.


Chromatin IP

Chromatin IP

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.

Chromatin IP

Chromatin IP

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.

Flow Cytometry

Flow Cytometry

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.


IF-IC

IF-IC

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).

Description

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.

Specificity / Sensitivity

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.

Source / Purification

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.

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).

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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

Toll Free: 1-800-387-1095

Fax: 1-800-563-3789

info.ca@neb.com

orders.ca@neb.com

About NEBTerms of Sale Web Discount Shipping Information Web Site Disclaimer Privacy Policy NEB USA Cell Signaling Technology Sitemap
Contents ¬©New England Biolabs Ltd.  New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.  New England Biolabs, Inc. is an ISO 9001 certified company.

Search another product:

 

Item has been added to the cart

 

Item has been added to the favourites