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Anti-dimethyl-Histone H3 (Lys27), clone 614M Antibody
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REFERENCES
Regulation of transcription by a protein methyltransferase.
Chen, D, et al. (1999) Science, 284: 2174-7 (1999)
Establishment of histone h3 methylation on the inactive X chromosome requires transient recruitment of Eed-Enx1 polycomb group complexes.
Silva, Jose, et al. (2003) Dev. Cell, 4: 481-95 (2003)
Species Reactivity Key Applications Host Format Antibody Type
Vrt WB, PIA Rabbit Culture Supernatant Monoclonal Antibody
Description:
Anti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M, rabbit monoclonal
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Specificity:
Histone H3 dimethylated on Lys27. An unidentified doublet sometimes appears by Western blotting in some lysates at ~35 kDa.
Molecular Weight:
17 kDa
Immunogen:
Peptide containing the sequence (ARme2KSA) in which me2 corresponds to dimethyl lysine at residue 27 of human histone H3.
Modifications:
Methylation
Clone:
614M
Isotype:
IgG
Background Information:
Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome. Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Histone H3 is methylated at Lys27 by EZH2, and and overexpression of EZH2 has been associated with both breast and prostate cancers. Methylation of H3K27 is involved in X chroosome inactivation, imprinting, circadian rhythms, and stem cell maintenance. H3K27me2 is a marker of classical heterochromatin.
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Species Reactivity:
Vertebrates
Species Reactivity Note:
Broad species cross-reactivity expected, based on sequence identity in most species.
Application Notes:
Western Blot Analysis:
1:1000-1:5000 dilutions of a previous lot of this clone detected dimethyl-Histone H3 in acid extracted proteins from HeLa cells, but did not detect unmethylated recombinant Histone H3 (Catalog # 14-494).
Peptide Inhibition:
0.5-2 µM of histone H3 peptides containing dimethyl-Lys27 abolished detection of histone H3 by anti-dimethyl-Histone H3 (Lys27), clone 614M (1:1000 dilution) in immunoblots of acid extracted proteins from HeLa cells.
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Control:
Acid extracted HeLa cell proteins
Quality Assurance:
Routinely evaluated by western blot in acid extrated protein from mitotic HeLa cells.
Western Blot Analysis:
1:500-2000 dilutions of this lot detected dimethyl-Histone H3 in acid extracted proteins from HeLa cells.
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Purification Method:
Unpurified supernatant
Presentation:
Cultured supernantant in 0.05% sodium azide.
Storage Conditions:
1 year at -20°C from date of shipment
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/ th |