Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP964Y] to Histone H3 (acetyl K14) - ChIP Grade
  • Suitable for: ChIP, WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EP964Y] to Histone H3 (acetyl K14) - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    There 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 Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    IHC-P
    Rat
    Human
    WB
    Rat
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (acetyl K14). The exact sequence is proprietary.
    (Peptide available as ab207713)

  • Positive control

    • WB: C6, Hek293T and HEL cell lysates. IHC-P: Human uterus adenocarcinoma and endometrium carcinoma tissues, Mouse kidney tissue and Rat liver tissue. ICC/IF: HeLa cells treated/untreated with trichostatin A. ChIP: Chromatin was prepared from HeLa cells (treated with 50 ng/ml nocodazole for 14 hours).

  • General notes

    Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

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

    Chromatin was prepared from HeLa cells (treated with 50 ng/ml nocodazole for 14 hours) according to the Abcam X-ChIP protocol. Cells were fixed with EGS (1.5 mM) for 30 minutes then formaldehyde (1%) for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab52946 (red), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (TaqMan approach).

  • Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    All lanes : Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor glial cell) treated with Trichostatin A whole cell lysates
    Lane 2 : 293T (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 3 : HEL (Human Erythroleukemia erythroblast) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 11 kDa
    Observed band size: 15 kDa
    why is the actual band size different from the predicted?


    Exposure time: 60 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST
     

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

    Immunocytochemistry/Immunofluorescence analysis of untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and TSA (Trichostatin A) (500ng/ml, 4h) treated HeLa cells labeling Histone H3 (acetyl K14) with purified ab52946 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100, counterstained with ab150120 AlexaFluor®594 Goat anti-Mouse secondary 1:1000  (2ug/ml). An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Negative Control 1: Rabbit primary antibody and anti-mouse secondary antibody(ab150120)

    Negative Control 2: Mouse primary antibody(ab7291) and anti-rabbit secondary antibody(ab150077)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

    Immunohistochemical analysis of paraffin-embedded Human endometrium carcinoma tissue labeling Histone H3 with ab52946, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on human endometrium carcinoma. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

    Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Histone H3 with ab52946, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on rat liver. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

    Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Histone H3 with ab52946, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on mouse kidney. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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

    Lane 1 : C6 (Rat glial tumor cell line) cell lysates untreated
    Lane 2 : C6 cell lysates treated with Trichostatin A

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 11 kDa
    Observed band size: 11 kDa

  • Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946) Anderson et al PLoS Negl Trop Dis. 2017 Apr 13;11(4):e0005539. doi: 10.1371/journal.pntd.0005539. eCollection 2017 Apr. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Histone H3 post-translational modifications profile in S. mansoni under HDAC inhibitor effect.

    Western blotting of the enriched histone protein fractions extracted from S. mansoni adult worms is shown for three biological replicates. Each biological replicate consisted of worms treated for 24 h with 1 μM TSA (lanes 2, 4 and 6) or with vehicle (lanes 1, 3 and 5).

    Additionally, antibody anti-H3 was used as a sample loading normalizer.

    Molecular weight marker ladder (L) is indicated.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

    ab52946 at 1/100 dilution staining Histone H3 (acetyl K14) in human uterus adenocarcinoma tissue by Immunohistochemistry, Paraffin embedded tissue.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

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