Recombinant
RabMAb

Recombinant Anti-Histone H3 (phospho S28) antibody [EP1874Y] (ab76312)

Overview

  • Product name

    Anti-Histone H3 (phospho S28) antibody [EP1874Y]
    See all Histone H3 primary antibodies
  • Description

    Rabbit monoclonal [EP1874Y] to Histone H3 (phospho S28)
  • Host species

    Rabbit
  • Specificity

    This antibody detects Histone H3 and Histone H3.3 when phosphorylated on Serine 28. It does not detect H3.3 when phosphorylated on Serine 31.
  • Tested applications

    Suitable for: WB, Dot blotmore details
    Unsuitable for: Flow Cyt,ICC,IHC-P or IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Chicken
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (phospho S28).
    Database link: P84243

  • Positive control

    • HeLa cell lysates treated with FBS + calyculin A.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76312 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
WB 1/2000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
Dot blot 1/100.
  • Application notes
    Is unsuitable for Flow Cyt,ICC,IHC-P or IP.
  • 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

    • All lanes : Anti-Histone H3 (phospho S28) antibody [EP1874Y] (ab76312) at 1/5000 dilution

      Lane 1 : HeLa cell lysates untreated
      Lane 2 : HeLa cell lysates treated with FBS + calyculin A

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 15 kDa
      Observed band size: 17 kDa
      why is the actual band size different from the predicted?

    • Dot blot performed using ab76312 at a dilution of 1/100. Lane 1 - Unmodified H3 peptide. Lane 2 - H3S28ph peptide. Lane 3 - H3.3S28ph peptide. Lane 4 - H3.3S31ph peptide. A HRP-conjugated goat anti-rabbit (H+L) was used as the secondary antibody at a dilution of 1/1000. The exposure time was 3 minutes and the dilution and blocking buffer used were 5% NFDM/TBST.

    References

    ab76312 has not yet been referenced specifically in any publications.

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