Recombinant
RabMAb

Recombinant Anti-Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) antibody [EPR18087] (ab188294)

Overview

  • Product name

    Anti-Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) antibody [EPR18087]
    See all Histone H1.3+Histone H1.4 primary antibodies
  • Description

    Rabbit monoclonal [EPR18087] to Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: PepArr, IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human Histone H1.4 aa 1-100 (phospho T17). The exact sequence is proprietary.
    Database link: P10412

  • Positive control

    • WB: HeLa treated with Colcemid acid extraction lysates; NIH/3T3 treated with 1.5µg /ml Colcemid for 12 hours whole cell lysates. IHC-P: Human, mouse and rat colon 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 ab188294 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.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/8000.
WB 1/5000. Detects a band of approximately 30 kDa (predicted molecular weight: 22 kDa).

Target

Images

  • All lanes : Anti-Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) antibody [EPR18087] (ab188294) at 1/5000 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 Colcemid acid extraction lysates

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 22 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) antibody [EPR18087] (ab188294) at 1/5000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) 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

    Predicted band size: 22 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • ab188294 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 H1.3 (phospho T17) + Histone H1.4 (phospho T17) with ab188294 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nucleus staining on epithelial cells of Human colon 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 colon tissue labeling Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) with ab188294 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nucleus staining on epithelial cells of mouse colon 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 H1.3 (phospho T17) + Histone H1.4 (phospho T17) with ab188294 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nucleus staining on epithelial cells of rat colon 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H1.3 (phospho T17) + Histone H1.4 (phospho T17) with ab188294 at 1/8000 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 ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab188294 at 1/8000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

References

This product has been referenced in:

  • Krishnan S  et al. Phospho-H1 Decorates the Inter-chromatid Axis and Is Evicted along with Shugoshin by SET during Mitosis. Mol Cell 67:579-593.e6 (2017). Read more (PubMed: 28781233) »
See 1 Publication for this product

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