Recombinant
RabMAb

Recombinant Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)

Overview

  • Product name

    Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184]
    See all Histone H2A (phospho S1) + Histone H4 (phospho S1) primary antibodies
  • Description

    Rabbit monoclonal [EPR18184] to Histone H2A (phospho S1) + Histone H4 (phospho S1)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: PepArr, Dot blot, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Histone H2A aa 1-100 (phospho S1). The exact sequence is proprietary.
    Database link: P04908

  • Positive control

    • WB: HeLa treated with 1.5µg/ml Colcemid for 12 hours whole cell lysates. IHC-P: human colon, mouse stomach, rat colon, human cerebral cortex, mouse heart and rat brain tissues. ICC/IF: HeLa cells.
  • 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 ab177309 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
PepArr Use at an assay dependent concentration.
Dot blot 1/1000.
WB 1/500. Detects a band of approximately 11, 14 kDa (predicted molecular weight: 11, 14 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/4000.

Target

Images

  • All lanes : Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309) at 1/500 dilution

    Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 1.5µg/ml Colcemid for 12 hours whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 11, 14 kDa
    Observed band size: 11,14 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Dot blot analysis of Histone H2A (phospho S1) peptide (Lane 1), unmodified Histone H2A peptide (Lane 2), Histone H4 (phospho S1) peptide (Lane 3), and unmodified Histone H4 peptide (Lane 4), labeled using ab177309 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab177309 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

  • ab177309 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
     
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
     
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on human colon tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on mouse stomach tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on rat colon tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on neuron cells of human cerebral cortex is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on mouse heart tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on neuron cells of rat brain tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

ab177309 has not yet been referenced specifically in any publications.

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