RabMAb

Anti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP Grade (ab52946)

Overview

  • Product nameAnti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Rabbit monoclonal [EP964Y] to Histone H3 (acetyl K14) - ChIP Grade
  • SpecificityThere was no cross-reactivity observed with recombinant H3 or the following modifications Acetyl-K9/pS10, -K18, -K23, and -K27 in dot plot. We have data to indicate that this antibody will not cross react with mouse. The antibody was tested against brain, kidney, spleen and heart lysates.
  • Tested applicationsIHC - Wholemount, ChIP, WB, IP, IHC-P, ICC/IF more details
  • Species reactivity
    Reacts with: Rat, Human, Saccharomyces cerevisiae, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe

    Does not react with

    Mouse
  • Immunogen

    A synthetic acetyl-specific peptide corresponding to residues around Lysine 14 of Histone H3

  • Positive control
    • C6 cell lysates, human adenocarcinoma of uterus tissue and HeLa cells.
  • General notes

    Produced under U.S. Patent No. 5,675,063. A 40 μl trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52946 in the following tested applications.

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

Application Notes
IHC - Wholemount Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. PubMed: 19188451
WB 1/2000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
IP 1/20.
IHC-P 1/100 - 1/250.
ICC/IF 1/250 - 1/500.
  • Application notesIs unsuitable for Flow Cyt.
  • 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
    • Alternative names
      • H 3 antibody
      • H3 antibody
      • H3/A antibody
      • H31_HUMAN antibody
      • H3FA antibody
      • H4F antibody
      • HIST1H3J antibody
      • HIST1H4 antibody
      • Histone cluster 1, H3a antibody
      • Histone H3.1 antibody
      • Histone H3.3 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
      • Histone H4 antibody
      see all

    Anti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP Grade images

    • All lanes : Anti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP Grade (ab52946) at 1/2000 dilution

      Lane 1 : C6 cell lysates untreated
      Lane 2 : C6 cell lysates treated with TSA

      Lysates/proteins at 10 µg per lane.

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa
    • ab52946 at 1/100-1/250 dilution staining Histone H3 (acetyl K14) in human adenocarcinoma of uterus by Immunohistochemistry, Paraffin embedded tissue.ab52946 at 1/100-1/250 dilution staining Histone H3 (acetyl K14) in human adenocarcinoma of uterus by Immunohistochemistry, Paraffin embedded tissue.
    • ab52946 at 1/250-1/500 dilution staining Histone H3 (acetyl K14) in HeLa cells by Immunofluorescence.
    • ICC/IF image of ab52946 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52946, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • IHC - Wholemount of Caenorhabditis elegans embryo labelling Histone H3 with ab52946. Sample was incubated with primary antibody (1/200 in TBS + 5% BSA + 0.1% Triton-X 100) for 12 hours at 4°C. ab150077, an Alexa Flour® 488-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • ab52946 immunoprecipitating Histone H3 (acetyl K14) in Drosophila melanogaster embryo whole tissue lysate. 500µg of tissue lysate was incubated with the undiluted primary antibody and matrix (Protein G) for 2 hours at 4°C. For western blotting ab52946 (1/20000) was used to confirm successful immunoprecipation.

      See Abreview

    References for Anti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP Grade (ab52946)

    This product has been referenced in:
    • Evellin S  et al. FOSL1 controls the assembly of endothelial cells into capillary tubes by direct repression of av and ß3 integrin transcription. Mol Cell Biol 33:1198-209 (2013). Human . Read more (PubMed: 23319049) »
    • Khan DH  et al. RNA-dependent dynamic histone acetylation regulates MCL1 alternative splicing. Nucleic Acids Res N/A:N/A (2013). Read more (PubMed: 24234443) »

    See all 10 Publications for this product

    Product Wall

    Application Immunoprecipitation
    Immuno-precipitation step Protein G
    Sample Caenorhabditis elegans Tissue lysate - whole (C. elegans whole body)
    Specification C. elegans whole body
    Total protein in input 800 µg
    Username

    Abcam user community

    Verified customer

    Submitted Mar 20 2014

    Application Immunoprecipitation
    Immuno-precipitation step Protein G
    Sample Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Fruit fly embryo)
    Specification Fruit fly embryo
    Total protein in input 500 µg
    Username

    Abcam user community

    Verified customer

    Submitted Mar 20 2014

    Application Immunoprecipitation
    Immuno-precipitation step Protein G
    Sample Mouse Cell lysate - nuclear (Nuclear extract of mouse embryonic fibroblasts)
    Specification Nuclear extract of mouse embryonic fibroblasts
    Total protein in input 200 µg
    Username

    Abcam user community

    Verified customer

    Submitted Mar 20 2014

    Application IHC - Wholemount
    Sample Caenorhabditis elegans Embryo (C. elegans embryo)
    Specification C. elegans embryo
    Username

    Abcam user community

    Verified customer

    Submitted Mar 18 2014

    Application Flow Cytometry
    Fixation Paraformaldehyde
    Permeabilization Yes - eBioscience Permeabilization Buffer
    Sample Human Cell (Differentiated Haematopoietic Stem Cells)
    Specification Differentiated Haematopoietic Stem Cells
    Gating Strategy Isotype negative control (white)
    Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
    Sample buffer: PBS and 10% FBS
    Username

    Abcam user community

    Verified customer

    Submitted Feb 24 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 23°C
    Sample Mouse Cell (Mouse embryonic fibroblasts)
    Specification Mouse embryonic fibroblasts
    Permeabilization Yes - 0.3% Triton X-100
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Nov 28 2013

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (12)
    Sample Human Cell lysate - nuclear (Hela cell)
    Specification Hela cell
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 11 2013

    Thank you for your response.

    My colleague Padamjeet is out of office this afternoon. I have checked the batch number you have provided (374056 )but unfortunatelyit seems our system does not recognize it. Could you please check you record agai...

    Read More

    Thank you for contacting us. Your inquiry has been forwarded to me by my colleague Davide.

    In order to understand the problem could you provide more information about the following questions;

    - The antibody ab45166 is indeed specific...

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    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.5% Triton-X100 in PBS
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Apr 18 2012

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