Recombinant
RabMAb

Recombinant Anti-Histone H3 (acetyl K27) antibody [EP865Y] - ChIP Grade (ab45173)

Overview

  • Product name
    Anti-Histone H3 (acetyl K27) antibody [EP865Y] - ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit monoclonal [EP865Y] to Histone H3 (acetyl K27) - ChIP Grade
  • Host species
    Rabbit
  • Specificity

    There is cross-reactivity with H2BK5Ac (Histone H2B acetylated on Lys 5).

  • Tested applications
    Suitable for: ChIP, WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (acetyl K27). The exact sequence is proprietary.

  • Positive control
    • WB: C6 cell lysate, treated with TSA. ICC/IF: A431 cells. IHC-P: Human hepatocellular carcinoma.
  • General notes

     

     

    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 ab45173 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
ChIP Use at an assay dependent concentration.
WB 1/500. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt 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

    • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of  ab45173 (blue), and 20µl of A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (TaqMan approach).

    • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of  ab45173 (blue), and 20µl of A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).

    • All lanes : Anti-Histone H3 (acetyl K27) antibody [EP865Y] - ChIP Grade (ab45173) at 1/500 dilution

      Lane 1 : C6 cell lysate, untreated
      Lane 2 : C6 cell lysate, treated with TSA

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP labeled at 1/2000 dilution

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

    • ab45173 (1/100) staining human Histone H3T in human hepatocellular carcinoma by immunohistochemistry using paraffin embedded tissue.

    • Immunofluorescent staining of A431 cells using ab45173 (1/100).

    References

    This product has been referenced in:
    • Song Y  et al. Long non-coding RNA HOTAIR mediates the switching of histone H3 lysine 27 acetylation to methylation to promote epithelial-to-mesenchymal transition in gastric cancer. Int J Oncol 54:77-86 (2019). Read more (PubMed: 30431069) »
    • Janssens DH  et al. Automated in situ chromatin profiling efficiently resolves cell types and gene regulatory programs. Epigenetics Chromatin 11:74 (2018). Read more (PubMed: 30577869) »
    See all 10 Publications for this product

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