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    human-histone-h3-phospho-s10-peptide-ab11477.pdf

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Epigenetics and Nuclear Signaling Histones H3 Phosphorylated
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Human Histone H3 (phospho S10) peptide (ab11477)

  • Datasheet
  • SDS
Reviews (2)Q&A (1)References (3)

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Western blot - Human Histone H3 (phospho S10) peptide (ab11477)

    Key features and details

    • Suitable for: Blocking

    Description

    • Product name

      Human Histone H3 (phospho S10) peptide
      See all Histone H3 proteins and peptides
    • Accession

      P68431
    • Animal free

      No
    • Nature

      Synthetic
      • Species

        Human
      • Modifications

        phospho S10

    Associated products

    • Corresponding Antibody

      • Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)
      • Anti-Histone H3 (phospho S10) antibody (ab5176)
    • Related Products

      • Human Histone H3 (unmodified ) peptide (ab2903)

    Specifications

    Our Abpromise guarantee covers the use of ab11477 in the following tested applications.

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

    • Applications

      Blocking - Blocking peptide for Anti-Histone H3 (phospho S10) antibody (ab5176), Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

    • Form

      Lyophilized
    • Additional notes

      - First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
      - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
      - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
      - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
      - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

    • Concentration information loading...

    Preparation and Storage

    • Stability and Storage

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

      Information available upon request.

    General Info

    • Alternative names

      • H3 histone family member E pseudogene
      • H3 histone family, member A
      • H3/A
      • H31_HUMAN
      • H3F3
      • H3FA
      • Hist1h3a
      • HIST1H3B
      • HIST1H3C
      • HIST1H3D
      • HIST1H3E
      • HIST1H3F
      • HIST1H3G
      • HIST1H3H
      • HIST1H3I
      • HIST1H3J
      • HIST3H3
      • histone 1, H3a
      • Histone cluster 1, H3a
      • Histone H3 3 pseudogene
      • Histone H3.1
      • Histone H3/a
      • Histone H3/b
      • Histone H3/c
      • Histone H3/d
      • Histone H3/f
      • Histone H3/h
      • Histone H3/i
      • Histone H3/j
      • Histone H3/k
      • Histone H3/l
      see all
    • 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.
    • Target information above from: UniProt accession P68431 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt

    Images

    • Western blot - Human Histone H3 (phospho S10) peptide (ab11477)
      Western blot - Human Histone H3 (phospho S10) peptide (ab11477)
      Lysates: Lane 1,3,5: 0.5µg/lane untreated histone Lane 2,4,6: 0.5µg/lane colcemid treated histone - treated Primary antibody: Lane 1-2 : ab5176 (1?g/ml) Lane 3-4 :ab5176 (1?g/ml) + phospho peptide (ab5176) 1?g/ml Lane 5-6 :ab5176 (1?g/ml) + non-phospho peptide ( ab2903 batch 3) 1?g/ml

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (3)

    Publishing research using ab11477? Please let us know so that we can cite the reference in this datasheet.

    ab11477 has been referenced in 3 publications.

    • Khan DH  et al. Mitogen-induced distinct epialleles are phosphorylated at either H3S10 or H3S28, depending on H3K27 acetylation. Mol Biol Cell 28:817-824 (2017). PubMed: 28077620
    • Drobic B  et al. Promoter chromatin remodeling of immediate-early genes is mediated through H3 phosphorylation at either serine 28 or 10 by the MSK1 multi-protein complex. Nucleic Acids Res 38:3196-208 (2010). Blocking . PubMed: 20129940
    • McManus KJ & Hendzel MJ The relationship between histone H3 phosphorylation and acetylation throughout the mammalian cell cycle. Biochem Cell Biol 84:640-57 (2006). Competitive Binding Assays . PubMed: 16936834

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews

    Other (Dot Blot) Abreview for Histone H3 peptide - phospho S10

    Excellent
    Abreviews
    Abreviews
    Application
    Other - Do not use

    Review text: I used this product to do a dot blot specificity test for cross reactivity of modified histone antibodies and everything worked great. I made two dilutions of the peptide and spotted it on a nitrocellulose membrane using a Bio-Dot apparatus. Once the peptides were bound to the membrane I incubated with the appropriate primary and secondary antibodies at an assay dependent concentration and developed using ECL.

    No primary antibody directed towards H3S10phos was used in this dot blot assay. This peptide was chosen to ensure there was no cross reactivity between the antibodies we are using on a regular basis. We have yet to see any cross reactivity with this peptide using 12+ antibodies directed towards histone H3 modified antibodies (some from Abcam some not).
    Sample: Human Purified protein

    Primary antibody (in addition to 'Histone H3 peptide - phospho S10')
    Primary antibody: None used

    Secondary antibody
    Secondary antibody: None used

    Abcam user community

    Verified customer

    Submitted Nov 01 2010

    Other (Dot Blot) Abreview for Histone H3 peptide - phospho S10

    Excellent
    Abreviews
    Abreviews
    Application
    Other - Do not use

    Review text: I used this product to do a dot blot specificity test for cross reactivity of modified histone antibodies and everything worked great. I made two dilutions of the peptide and spotted it on a nitrocellulose membrane using a Bio-Dot apparatus. Once the peptides were bound to the membrane I incubated with the appropriate primary and secondary antibodies at an assay dependent concentration and developed using ECL.
    Sample: Human Purified protein

    Primary antibody (in addition to 'Histone H3 peptide - phospho S10')
    Name: Non-Abcam antibody was used: anti-H2A.Z
    Host species: Mouse
    Clonality: Polyclonal
    Dilution: 1/5000

    Secondary antibody
    Name: Non-Abcam antibody was used: anti-rabbit
    Host species: Mouse
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/10000

    Abcam user community

    Verified customer

    Submitted Oct 30 2010

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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