Buy Three, Get the Fourth Free! | Start Saving >>
53312
Methyl-Histone H3 (Lys27) Antibody Sampler Kit
Primary Antibodies

Methyl-Histone H3 (Lys27) Antibody Sampler Kit #53312

Chromatin IP-seq Image 1

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Ezh2 (D2C9) XP® Rabbit mAb #5246 or Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. EZH2 and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both EZH2 and H3K27me3 across MYT1, a known target gene of H3K27me3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Learn more about how we get our images
Western Blotting Image 2

Western blot analysis of various cell lines using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 3

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 4

Western blot analysis of extracts from HeLa and C2C12 cell lines using Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 5

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

Learn more about how we get our images
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.

Learn more about how we get our images
IHC-P (paraffin) Image 7

Immunohistochemical analysis of paraffin-embedded human lymphoma using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb in the presence of non-methyl peptide (left) or K27 tri-methyl peptide (right).

Learn more about how we get our images
Chromatin IP Image 8

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys27) (C36B11) 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 Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human MYT-1 Exon 1 Primers #4493. 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.

Learn more about how we get our images
Flow Cytometry Image 9

Flow cytometric analysis of human peripheral blood lymphocytes using Di-Methyl-Histone H3 (Lys27) (D18C8) 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.

Learn more about how we get our images
IP Image 10

Immunoprecipitation of mono-methyl-histone H3 Lys27 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb. Western blot analysis was performed using Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 11

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

Learn more about how we get our images
Flow Cytometry Image 12

Flow cytometric analysis of human whole blood cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (blue) and 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. Analysis was performed on cells in the lymphocyte gate.

Learn more about how we get our images
IF-IC Image 13

Confocal immumunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb (green). Actin filaments have been labled with DY-554 phalloidin (red).

Learn more about how we get our images
Flow Cytometry Image 14

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.

Learn more about how we get our images
IF-IC Image 15

Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Learn more about how we get our images
Chromatin IP Image 16

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Di-Methyl-Histone H3 (Lys27) (D18C8) 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 GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human AFM Intron 1 Primers #5098. 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.

Learn more about how we get our images
IF-IC Image 17

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).

Learn more about how we get our images
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb 9733 20 µl
  • WB
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb 9728 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb 84932 20 µl
  • WB
  • IP
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 (Lys27) Antibody Sampler Kit provides an economical means of detecting levels of mono-, di-, and tri-methyl histone H3 Lys27 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 (Lys27) Antibody Sampler Kit detects endogenous levels of its target protein. Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb detects endogenous levels of histone H3 only when tri-methylated on Lys27. The antibody does not cross-react with non-methylated, mono-methylated or di-methylated Lys27. Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb detects endogenous levels of histone H3 when di-methylated on Lys27. The antibody does show some cross-reactivity with mono-methylated Lys27, but does not cross-react with non-methylated or tri-methylated Lys27. Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb recognizes endogenous levels of histone H3 protein only when mono-methylated at Lys27. This antibody does not cross-react with non-methylated, di-methylated, or tri-methylated Lys27. 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.

Monoclonal methyl-histone H3 Lys27 antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of histone H3 in which Lys27 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 Lys27 is generally associated with transcriptional repression and regulation of stem cell pluripotency and differentiation, through the regulation of facultative heterochromatin. Mono-, di-, and tri-methylation of histone H3 Lys27 are all mediated by polycomb repressor complex 2 (PRC2), which contains either the EZH1 or EZH2 methyltransferase proteins. Tri- and di-methyl-histone H3 Lys27 levels are highest at the promoters of polycomb-repressed genes. In addition, tri- and di-methyl-histone H3 Lys27 is found at the promoters of inactive, but transcriptionally poised genes that also contain the active tri-methyl-histone H3 Lys4 modification. Mono-methyl-histone H3 Lys27 is more widely dispersed and found in the bodies of active genes.

  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.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Tween is a registered trademark of ICI Americas, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.