Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)


  • Product nameAnti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit monoclonal [Y49] to Histone H3 (di methyl K9) - ChIP Grade
  • SpecificityThe antibody only detects Histone H3 dimethylated on Lysine 9.
  • Tested applicationsICC/IF, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (di methyl K9).

  • Positive control
    • WB: HeLa cell lysate. ICC/IF: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.


    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.



Our Abpromise guarantee covers the use of ab32521 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500.
WB 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
ChIP Use 5 µg for 25 µg of chromatin.
  • Application notesIs unsuitable for Flow Cyt,IHC or IP.
  • Target

    • FunctionCore 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.
    • Sequence similaritiesBelongs to the histone H3 family.
    • Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
    • Post-translational
      Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
      Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
      Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
      Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
      Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) 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 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
      Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). 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.
    • Cellular localizationNucleus. Chromosome.
    • Information by UniProt
    • Database links
    • Alternative names
      • H3 histone family, member J antibody
      • FLJ92264 antibody
      • H3 histone family, member A antibody
      • H3 histone family, member B antibody
      • H3 histone family, member C antibody
      • H3 histone family, member D antibody
      • H3 histone family, member F antibody
      • H3 histone family, member H antibody
      • H3 histone family, member I antibody
      • H3 histone family, member K antibody
      • H3 histone family, member L antibody
      • H3 histone, family 3A antibody
      • H3.3A antibody
      • H3/a antibody
      • H3/b antibody
      • H3/c antibody
      • H3/d antibody
      • h3/f antibody
      • H3/h antibody
      • H3/i antibody
      • H3/j antibody
      • H3/k antibody
      • H3/l antibody
      • H31_HUMAN antibody
      • H3F1K antibody
      • H3F3 antibody
      • H3FA antibody
      • H3FB antibody
      • H3FC antibody
      • H3FD antibody
      • H3FF antibody
      • H3FH antibody
      • H3FI antibody
      • H3FJ antibody
      • H3FK antibody
      • H3FL antibody
      • HIST1H3A antibody
      • HIST1H3B antibody
      • HIST1H3C antibody
      • HIST1H3D antibody
      • HIST1H3E antibody
      • HIST1H3F antibody
      • HIST1H3G antibody
      • HIST1H3H antibody
      • HIST1H3I antibody
      • HIST1H3J antibody
      • HIST3H3 antibody
      • Histone 1, H3a antibody
      • Histone 1, H3b antibody
      • Histone 1, H3c antibody
      • Histone 1, H3d antibody
      • Histone 1, H3e antibody
      • Histone 1, H3f antibody
      • Histone 1, H3g antibody
      • Histone 1, H3h antibody
      • Histone 1, H3i antibody
      • Histone 1, H3j antibody
      • Histone cluster 1, H3a antibody
      • Histone cluster 1, H3b antibody
      • Histone cluster 1, H3c antibody
      • Histone cluster 1, H3d antibody
      • Histone cluster 1, H3e antibody
      • Histone cluster 1, H3f antibody
      • Histone cluster 1, H3g antibody
      • Histone cluster 1, H3i antibody
      • Histone cluster 1, H3j antibody
      • Histone H 3 antibody
      • Histone H3.1 antibody
      • Histone H3.2 antibody
      • Histone H3/a antibody
      • Histone H3/b antibody
      • Histone H3/c antibody
      • Histone H3/d antibody
      • Histone H3/f antibody
      • Histone H3/h antibody
      • Histone H3/i antibody
      • Histone H3/j antibody
      • Histone H3/k antibody
      • Histone H3/l antibody
      • Histone H3/m antibody
      • Histone H3/o antibody
      see all

    Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade images

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Histone H3 (di methyl K9) with purified ab32521 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

      Control: PBS only

    • All lanes : Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : recombinant Histone H3

      Predicted band size : 15 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)
    • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32521 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

    • ab32521 (1/500) staining Histone H3 di-methyl K9 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details, please refer to Abreview.

      See Abreview

    References for Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)

    This product has been referenced in:
    • Dalvoy Vasudevarao M  et al. Inhibition of acetyltransferase alters different histone modifications: probed by small molecule inhibitor plumbagin. J Biochem 152:453-62 (2012). Read more (PubMed: 22923743) »

    See 1 Publication for this product

    Product Wall

    For detecting nuclear proteins via western blot we recommend using a RIPA buffer or cell fractionation protocol in preparation of your cell lysate. I attached our protocol book to this email for information regarding specific cell lysate preparation pr...

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    All of our products are for research purposes only and are not certified for clinical research applications.

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.5% Triton X100

    Dr. Kirk McManus

    Verified customer

    Submitted Sep 07 2011