04-821 | Anti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M, rabbit monoclonal

04-821
100 µL  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      VrtWB, PIARbCulture SupernatantMonoclonal Antibody
      Description
      Catalogue Number04-821
      DescriptionAnti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M, rabbit monoclonal
      Alternate Names
      • H3K27me2
      • Histone H3 (di methyl K27)
      • H3 histone family, member M
      • H3 histone, family 2
      • histone 2
      • H3c
      • histone cluster 2
      • H3c
      Background InformationHistones 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.
      References
      Product Information
      FormatCulture Supernatant
      Control
      • Acid extracted HeLa cell proteins
      PresentationCultured supernantant in 0.05% sodium azide.
      Applications
      ApplicationAnti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys27) also known as H3K27me2, Histone H3 (di methyl K27) & has been validated in WB, PIA.
      Key Applications
      • Western Blotting
      • Peptide Inhibition Assay
      Application NotesWestern 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.
      Biological Information
      ImmunogenPeptide containing the sequence (ARme2KSA) in which me2 corresponds to dimethyl lysine at residue 27 of human histone H3.
      Clone614M
      HostRabbit
      SpecificityHistone H3 dimethylated on Lys27. An unidentified doublet sometimes appears by Western blotting in some lysates at ~35 kDa.
      IsotypeIgG
      Species Reactivity
      • Vertebrates
      Species Reactivity NoteBroad species cross-reactivity expected, based on sequence identity in most species.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in a histone cluster on chromosome 1. This gene is one of four histone genes in the cluster that are duplicated; this record represents the telomeric copy. [provided by RefSeq]
      Gene Symbol
      • H3
      • H3.2
      • H3/M
      • H3/m
      • H3/o
      • H3F2
      • H3FM
      • MGC9629
      • OTTHUMP00000014041
      Modifications
      • Methylation
      Purification MethodUnpurified supernatant
      UniProt Number
      UniProt SummaryFUNCTION:Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. During nucleosome assembly the chaperone ASF1A interacts with the histone H3-H4 heterodimer.

      SUBCELLULAR LOCATION: Nucleus.

      Developmental stage Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

      PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18.

      Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription.

      Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression.

      Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin.

      Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase, probably by DAPK3. Phosphorylation at 'Ser-11' by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation.

      Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.

      SIMILARITY:Belongs to the histone H3 family.

      Molecular Weight17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely 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.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage Conditions1 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/ thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-dimethyl-Histone H3 (Lys27), clone 614M - 2301273 2301273
      Anti-dimethyl-Histone H3 (Lys27), clone 614M - 2309044 2309044
      Anti-dimethyl-Histone H3 (Lys27), clone 614M - 2829136 2829136
      Anti-dimethyl-Histone H3 (Lys27), clone 614M - DAM1551732 DAM1551732
      Anti-dimethyl-Histone H3 (Lys27), clone 614M Monoclonal Antibody 2947638

      References

      Reference overviewPub Med ID
      Establishment of histone h3 methylation on the inactive X chromosome requires transient recruitment of Eed-Enx1 polycomb group complexes.
      Silva, Jose, et al.
      Dev. Cell, 4: 481-95 (2003)  2003

      Show Abstract
      12689588 12689588
      Regulation of transcription by a protein methyltransferase.
      Chen, D, et al.
      Science, 284: 2174-7 (1999)  1999

      Show Abstract
      10381882 10381882

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      Categories

      Life Science Research > Antibodies and Assays > Primary Antibodies