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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.
Quality LevelMQ100
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
Global Trade Item Number
Catalogue Number GTIN
04-821 04053252472602

Documentation

Anti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M, rabbit monoclonal SDS

Title

Safety Data Sheet (SDS) 

Anti-dimethyl-Histone H3 (Lys27) Antibody, clone 614M, rabbit monoclonal 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 - 3224214 3224214
Anti-dimethyl-Histone H3 (Lys27), clone 614M - 4083058 4083058
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