05-1249 | Anti-Dimethyl Histone H3 (Lys9) Antibody, clone CMA307

05-1249
100 µg  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, VrtWB, ELISA, ICC, IP, Mplex, ChIPMPurifiedMonoclonal Antibody
      Description
      Catalogue Number05-1249
      Replaces04-791
      DescriptionAnti-Dimethyl Histone H3 (Lys9) Antibody, clone CMA307
      Alternate Names
      • H3K4me2
      • Histone H3 (di methyl K4)
      • H3 histone family, member T
      • histone 3
      • H3
      • histone cluster 3
      • H3
      Background InformationHistone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. Histone proteins are highly post-translationally modified, and Histone H3 is the most extensively modified of the five histones. Histone H3 sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes. Di- and tri-methylation of histone H3 are gene silencing marks, while removal of these marks by the demethylase genes Jmjd1a and Jmjd2c leads to gene activation.
      References
      Product Information
      FormatPurified
      Control
      • HeLa acid extract
      PresentationMouse monoclonal IgG1κ supernatant in PBS with 0.05% sodium azide.
      Applications
      ApplicationUse Anti-Dimethyl Histone H3 (Lys9) Antibody, clone CMA307 (mouse monoclonal antibody) validated in WB, ELISA, ICC, IP, Mplex to detect Dimethyl Histone H3 (Lys9) also known as H3K9me2, Histone H3 (di methyl K9).
      Key Applications
      • Western Blotting
      • ELISA
      • Immunocytochemistry
      • Immunoprecipitation
      • Multiplexing
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation::
      Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG, or Anti-dimethyl Histone H3 (Lys9) antibody, clone CMA307 and the Magna ChIP G (Cat. # 17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers B-Globin.
      Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 05-1249, Anti-dimethyl H3 (Lys9), clone CMA307 at 2.0ug/mL (1:500) dilution. Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.

      ELISA:
      This antibody has been shown by an outside laboratory to be suitable for ELISA. Kimura, H., et al. (2008). Cell Struct. Funct. 33: 61-73.

      Immunocytochemistry:
      This antibody has been shown by an outside laboratory to be suitable for immunocytochemistry ( Kimura, 2008).

      Immunoprecipitation:
      This antibody has been shown by an outside laboratory to be suitable for immunoprecipitation ( Kimura, 2008).

      Multiplexing:
      This antibody specifically recognizes histone H3 dimethylated on Lys9 by Luminex® assay.
      Biological Information
      ImmunogenSynthetic peptide corresponding to amino acids 1-19 of human Histone H3, dimethylated on Lys9, conjugated to KLH.
      Epitopedimethyl Lys9
      CloneCMA307
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityThis antibody recognizes Histone H3 dimethylated at Lys9. Phosphorylation of Ser10 interferes with antibody binding to dimethyl Lys9.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Vertebrates
      Species Reactivity NoteHuman. Broad species cross-reactivity is expected, based on sequence homology.
      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. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone 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 located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq]
      Gene Symbol
      • H3.4
      • H3/g
      • H3/t
      • H3FT
      • H3T
      • H3t
      • MGC126886
      • MGC126888
      • OTTHUMP00000037945
      Modifications
      • Methylation
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q16695 # 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.
      SIZE: 136 amino acids; 15508 Da
      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.
      SUBCELLULAR LOCATION: Nucleus.
      TISSUE SPECIFICITY: Expressed in testicular cells.
      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 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & 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 (By similarity). & 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. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity).
      SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family.
      Molecular Weight~17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceWestern Blot Analysis: 1:10,000 of this lot detected Dimethyl Histone H3 in HeLa acid extract.
      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 ConditionsStable for 1 year at 2-8°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size100 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - 2137041 2137041
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - 2038281 2038281
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - 2040913 2040913
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - 2041595 2041595
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - JBC1765517 JBC1765517
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - NG1684656 NG1684656
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - NG1698980 NG1698980
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - NG1740980 NG1740980
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - NG1787245 NG1787245
      Anti-Dimethyl Histone H3 (Lys9), clone CMA307 - NG1800813 NG1800813

      References

      Reference overviewPub Med ID
      Transcriptional activation of transposable elements in mouse zygotes is independent of Tet3-mediated 5-methylcytosine oxidation.
      Inoue, A; Matoba, S; Zhang, Y
      Cell research  22  1640-9  2012

      Show Abstract
      23184059 23184059
      Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species.
      Lin, X; Tirichine, L; Bowler, C
      Plant methods  8  48  2012

      Show Abstract
      23217141 23217141
      Cooperative and antagonistic contributions of two heterochromatin proteins to transcriptional regulation of the Drosophila sex determination decision.
      Li, H; Rodriguez, J; Yoo, Y; Shareef, MM; Badugu, R; Horabin, JI; Kellum, R
      PLoS genetics  7  e1002122  2011

      Show Abstract
      21695246 21695246
      The organization of histone H3 modifications as revealed by a panel of specific monoclonal antibodies.
      Kimura, Hiroshi, et al.
      Cell Struct. Funct., 33: 61-73 (2008)  2008

      18227620 18227620
      Differential H3K4 methylation identifies developmentally poised hematopoietic genes.
      Orford, Keith, et al.
      Dev. Cell, 14: 798-809 (2008)  2008

      18477461 18477461

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      Life Science Research > Antibodies and Assays > Primary Antibodies