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RabMAbs (2134-1)

Anti-Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)

Submit an Abreview Q&A (1)Specific References (8)

Overview

  • Product nameAnti-Histone H3 (acetyl K56) antibody [EPR996Y]See all Histone H3 primary antibodies ...
  • Description
    Rabbit monoclonal [EPR996Y] to Histone H3 (acetyl K56)
  • Specificityab76307 only detects Histone H3 acetylated on Lysine 56.
  • Tested applicationsWB, IHC-P, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae
  • Immunogen

    A synthetic acetylated peptide corresponding to residues surrounding Lys 56 of Histone H3

  • Positive controlC6 cell lysate, untreated or treated with TSA; HeLa cells.
  • General notesProduced under U.S. Patent No. 5,675,063.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • PurityTissue culture supernatant
  • Clonality Monoclonal
  • Clone numberEPR996Y
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab76307 in the following tested applications.

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

Application Notes
WB
IHC-P
ICC/IF
  • Application notesICC/IF: 1/100 - 1/250.
    IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use of HRP-conjugated or polymerized HRP secondary antibodies, stronger signals have been found using the polymerized HRP secondary.
    WB: 1/1000 - 1/2000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).

    Is unsuitable for Flow Cyt or IP.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.

    • 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
      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 localizationNucleus. 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
    • Database links
      see all
    • Alternative names
        H 3 antibodyH3 antibodyH3/A antibody
        H31_HUMAN antibodyH3FA antibodyHIST1H3J antibodyHistone cluster 1, H3a antibodyHistone H3.1 antibodyHistone H3.3 antibodyHistone H3/a antibodyHistone H3/b antibodyHistone H3/c antibodyHistone H3/d antibodyHistone H3/f antibodyHistone H3/h antibodyHistone H3/i antibodyHistone H3/j antibodyHistone H3/k antibodyHistone H3/l antibody
      see all

    Anti-Histone H3 (acetyl K56) antibody [EPR996Y] images

    • Immunocytochemistry/ Immunofluorescence - Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      Immunocytochemistry/ Immunofluorescence - Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      ab76307 at 1/100 dilution staining Histone H3 in HeLa cells by Immunofluorescence.
    • Western blot - Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      Western blot - Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      All lanes : Anti-Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307) at 1/2000 dilution

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      goat anti-rabbit HRP at 1/1000 dilution

      Predicted band size : 15 kDa
      Observed band size : 15 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)
      IHC image of ab76307 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab76307, undiluted, for 15 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.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      ab76307 showing positive staining in Cervical carcinoma tissue.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      ab76307 showing positive staining in Gastric adenocarcinoma tissue.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      ab76307 showing positive staining in Lung adenocarcinoma tissue.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-Histone H3 (acetyl K56) antibody [EPR996Y](ab76307)
      ab76307 showing positive staining in Lymphoma tissue.

    References for Anti-Histone H3 (acetyl K56) antibody [EPR996Y] (ab76307)

    This product has been referenced in:
    • Tropberger P  et al. Regulation of transcription through acetylation of H3K122 on the lateral surface of the histone octamer. Cell 152:859-72 (2013). WB ; Human . Read more (PubMed: 23415232) »
    • Drogaris P  et al. Histone deacetylase inhibitors globally enhance h3/h4 tail acetylation without affecting h3 lysine 56 acetylation. Sci Rep 2:220 (2012). WB ; Human . Read more (PubMed: 22355734) »

    See all 8 Publications for this product

    Publishing research using ab76307 ? Please let us know so that we can cite the reference in this datasheet

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    What is the protocol used for IF with this antibody?

    Thank you for contacting Abcam regarding ab76307.


    Below please find the IF protocol we suggest:


    Solutions and reagents

    1.1. Washing buffer:
    PBST washing buffer: 1X PBS / 0.1% Tween-20


    1.2. 2% Parafor...

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