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05-1242 Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4

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      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, MPIA, IF, DB, ChIP-seq, WB, ChIPMPurifiedMonoclonal Antibody
      Catalogue Number05-1242
      DescriptionAnti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4
      Alternate Names
      • H3K9me3
      • Histone H3 (tri methyl K9)
      • H3 histone family, member T
      • histone 3
      • H3
      • histone cluster 3
      • H3
      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, the basic subunit of chromatin. Histone 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). Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely via acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α/β. The enzymes responsible for H3K9me3 formation are SUV39H1 and SUV39H2.
      Product Information
      • HeLa Acid Extract
      PresentationPurified antibody supplied at 1 mg/ml in 0.1 M Tris-glycine, pH 7.4, 150 mM NaCl, 0.05% NaN3.
      Quality LevelMQ100
      ApplicationUse Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 (mouse monoclonal antibody) validated in ChIP, PIA, IF, DB, ChIP-seq, WB to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).
      Key Applications
      • Peptide Inhibition Assay
      • Immunofluorescence
      • Dot Blot
      • ChIP-seq
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation (ChIP):
      Representative data from a previous lot. Sonicated 3T3 L1 chromatin was subjected to chromatin immunoprecipitation using anti- trimethyl-histone H3 (Lys9) and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of trimethylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using primers flanking the p16 promoter.

      Peptide Inhibition Analysis:
      Peptide blocking assay demonstrates distinct preference of the antibody for the trimethyl form vs. the dimethyl form.

      Chromatin Immunoprecipitation (ChIP):
      ChIP analysis of known chromosomal Suv39h targets (H3K9me3 in major satellites, mouseES cells).

      Dot Blot Analysis:
      Dot-blot analysis demonstrating specificity of anti-H3K9me3, clone 6F12-H4 for trimethyl Lys9 of histone H3.
      Biological Information
      ImmunogenKLH-conjugated, branched synthetic peptide containing the sequence (QTARme3K- STGGKA)2-KC, in which me3K corresponds to trimethyl lysine 9 of human histone H3. Data suggests this antibody may require the presence of the glutamine residue in position 5 for binding.
      EpitopeTrimethyl Lys9
      SpecificityRecognizes trimethyl-histone H3 (Lys9), Mr 17 kDa. Cross-reacts with dimethyl histone H3(Lys9) at higher concentrations.
      Species Reactivity
      • Human
      • Mouse
      Species Reactivity NoteHuman and mouse. Based on sequence homology, broad species cross-reactivity is expected with all mammals, Drosophila, Xenopus, and Arabidopsis.
      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
      • MGC126886
      • OTTHUMP00000037945
      • Methylation
      Purification MethodPurified
      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.


      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: Belongs to the histone H3 family.
      Molecular Weight~17 kDa
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by Western Blot on HeLa acid extracts.

      Western Blot Analysis: A 0.5 – 5 μg dilution of this lot detected trimethyl histone H3 (Lys9) in HeLa acid extracts.
      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 freezing, which can damage IgG and affect product performance.
      Packaging Information
      Material Size200 µL
      Transport Information
      Supplemental Information


      Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 SDS


      Safety Data Sheet (SDS) 

      Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 Certificates of Analysis

      TitleLot Number
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 2474943
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - NG1741706 NG1741706
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 2020467 2020467
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 2070421 2070421
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 2199715 2199715
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 2287194 2287194
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 2325757 2325757
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 3162441 3162441
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - 3275545 3275545
      Anti-trimethyl-Histone H3 (Lys9), clone 6F12-H4 - DAM160502 DAM160502


      Reference overviewApplicationPub Med ID
      Independent Mechanisms Target SMCHD1 to Trimethylated Histone H3 Lysine 9-Modified Chromatin and the Inactive X Chromosome.
      Brideau, NJ; Coker, H; Gendrel, AV; Siebert, CA; Bezstarosti, K; Demmers, J; Poot, RA; Nesterova, TB; Brockdorff, N
      Molecular and cellular biology  35  4053-68  2015

      Show Abstract
      26391951 26391951
      miR-142-5p and miR-130a-3p are regulated by IL-4 and IL-13 and control profibrogenic macrophage program.
      Su, S; Zhao, Q; He, C; Huang, D; Liu, J; Chen, F; Chen, J; Liao, JY; Cui, X; Zeng, Y; Yao, H; Su, F; Liu, Q; Jiang, S; Song, E
      Nature communications  6  8523  2015

      Show Abstract
      26436920 26436920
      Alterations of epigenetic signatures in hepatocyte nuclear factor 4α deficient mouse liver determined by improved ChIP-qPCR and (h)MeDIP-qPCR assays.
      Zhang, Q; Lei, X; Lu, H
      PloS one  9  e84925  2014

      Show Abstract
      24427299 24427299
      Ligand-dependent corepressor contributes to transcriptional repression by C2H2 zinc-finger transcription factor ZBRK1 through association with KRAB-associated protein-1.
      Calderon, MR; Verway, M; Benslama, RO; Birlea, M; Bouttier, M; Dimitrov, V; Mader, S; White, JH
      Nucleic acids research  42  7012-27  2014

      Show Abstract
      24829459 24829459
      Suv39h1 mediates AP-2α-dependent inhibition of C/EBPα expression during adipogenesis.
      Zhang, ZC; Liu, Y; Li, SF; Guo, L; Zhao, Y; Qian, SW; Wen, B; Tang, QQ; Li, X
      Molecular and cellular biology  34  2330-8  2014

      Show Abstract
      Western Blotting24732798 24732798
      High levels of glucose induce metabolic memory in cardiomyocyte via epigenetic histone H3 lysine 9 methylation.
      Xi-Yong Yu,Yong-Jian Geng,Jia-Liang Liang,Saidan Zhang,He-Ping Lei,Shi-Long Zhong,Qiu-Xiong Lin,Zhi-Xin Shan,Shu-Guang Lin,Yangxin Li
      Molecular biology reports  39  2012

      Show Abstract
      22707199 22707199
      p53-mediated heterochromatin reorganization regulates its cell fate decisions.
      Mungamuri, SK; Benson, EK; Wang, S; Gu, W; Lee, SW; Aaronson, SA
      Nature structural & molecular biology  19  478-84, S1  2012

      Show Abstract
      Western Blotting22466965 22466965
      Compensatory functions of histone deacetylase 1 (HDAC1) and HDAC2 regulate transcription and apoptosis during mouse oocyte development.
      Ma, P; Pan, H; Montgomery, RL; Olson, EN; Schultz, RM
      Proceedings of the National Academy of Sciences of the United States of America  109  E481-9  2012

      Show Abstract
      Immunofluorescence22223663 22223663
      Epigenomics: maternal high-fat diet exposure in utero disrupts peripheral circadian gene expression in nonhuman primates.
      Suter, M; Bocock, P; Showalter, L; Hu, M; Shope, C; McKnight, R; Grove, K; Lane, R; Aagaard-Tillery, K
      FASEB journal : official publication of the Federation of American Societies for Experimental Biology  25  714-26  2011

      Show Abstract
      21097519 21097519
      Corepressor protein CDYL functions as a molecular bridge between polycomb repressor complex 2 and repressive chromatin mark trimethylated histone lysine 27.
      Zhang, Y; Yang, X; Gui, B; Xie, G; Zhang, D; Shang, Y; Liang, J
      The Journal of biological chemistry  286  42414-25  2011

      Show Abstract
      22009739 22009739

      Technical Info

      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics (EMD)

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      Life Science Research > Antibodies and Assays > Primary Antibodies