Overview

  • Product name
    Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (tri methyl K9, phospho S10) - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ChIP, WB, ICC/IF, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (methyl K9, phospho S10) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab15644)

  • Positive control
    • HeLa Histone Preparation; Colcemid-treated HeLa histone Preparation; Hela whole cell extract

Properties

Applications

Our Abpromise guarantee covers the use of ab5819 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (tri methyl K9 or phospho S10) peptide (ab15644).
ICC/IF Use at an assay dependent concentration.
ICC 1/2000.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the histone H3 family.
  • Developmental stage
    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • Post-translational
    modifications
    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 localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H3 histone family member E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA 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 cluster 1, H3a antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.1 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
    see all

Images

  • Rabbit polyclonal to Histone H3 (phospho S10, tri methyl K9) - ab5819, on histone preparations from HeLa cells. Primary antibody used at 1/500.

    Lanes 1, 3, 5, 7, 9 - control HeLa histones (0.5ug/lane)
    Lanes 2, 4, 6, 8, 10 - Colcemid-treated HeLa histones (0.5ug/lane)

    All blocking peptides are 1ug/lane

    Lanes 1 and 2: ab5819  
    Lanes 3 and 4: ab5819 + Histone H3 (Tri Me K9, phospho S10) peptide - ab15644
    Lanes 5 and 6: ab5819 + Histone H3 peptide (unmodified) - ab7228
    Lanes 7 and 8: ab5819 + Histone H3  (phospho S10) peptide - ab11477
    Lanes 9 and 10: ab5819 + Histone H3 (tri methyl K9) peptide - ab1773 

    Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721 1/5000.

    ab5819 is specific for Histone H3 phosphorylated at residue S10 and tri methylated at residue K9. The activity of the antibody is blocked by the addition of the immunizing peptide, ab15644 (lanes 3 and 4).

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  • SKN cells stained with ab5819 (green) at a dilution of 1/2000. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab5819. The DNA is stained with DAPI (blue). 100x magnification.

  • ab5819 at a 1/500 dilution staining mitotic human HeLa cells by immunocytochemistry. The antibody was incubated with formaldehyde fixed cells for 1 1/2 hours and bound antibody was then detected using a Texas red conjugated goat anti-rabbit IgG.

    This image is courtesy of an Anonymous Abreview submitted on 6 April 2006.

    See Abreview

  • IHC image of Histone H3 (tri methyl K9, phospho S10) staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5819, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • References

    This product has been referenced in:
    • Rothbart SB  et al. An Interactive Database for the Assessment of Histone Antibody Specificity. Mol Cell 59:502-11 (2015). Read more (PubMed: 26212453) »
    • Sabbattini P  et al. An H3K9/S10 methyl-phospho switch modulates Polycomb and Pol II binding at repressed genes during differentiation. Mol Biol Cell 25:904-15 (2014). ChIP ; Mouse . Read more (PubMed: 24430871) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Formaldehyde
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%

    Abcam user community

    Verified customer

    Submitted Apr 06 2006

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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