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RecombinantRabMAb

Recombinant Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)

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ChIP - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Peptide Array - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
  • Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17589] to Histone H2A.X+Histone H2A (acetyl K5) - BSA and Azide free
  • Suitable for: PepArr, ChIP, WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free
    See all Histone H2A.X+Histone H2A primary antibodies

  • Description

    Rabbit monoclonal [EPR17589] to Histone H2A.X+Histone H2A (acetyl K5) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: PepArr, ChIP, WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: a wide range of other species

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • General notes

    ab250006 is the carrier-free version of ab177863.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab250006 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.
ChIP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 14,15 kDa (predicted molecular weight: 14,15 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
Notes
PepArr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 14,15 kDa (predicted molecular weight: 14,15 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.

Target

Images

  • ChIP - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    ChIP - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)

    This data was developed using ab177863, the same antibody clone in a different buffer formulation.

    Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab177863(red), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.
  • Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    All lanes : Anti-Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) antibody [EPR17589] - ChIP Grade (ab177863) at 1/4000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours. Whole cell lysates
    Lane 2 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500 ng/ml Trichostatin A for 4 hours. Whole cell lysates
    Lane 4 : Untreated NIH/3T3 (Human epithelial cell line from cervix adenocarcinoma) 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: 14,15 kDa
    Observed band size: 14,15 kDa why is the actual band size different from the predicted?



    This data was developed using ab177863, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)

    This data was developed using ab177863, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cell line labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. 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: ab177863 at 1/400 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.
  • Peptide Array - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Peptide Array - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    This data was developed using ab177863, the same antibody clone in a different buffer formulation.ab177863 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.
  • Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    All lanes : Anti-Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) antibody [EPR17589] - ChIP Grade (ab177863) at 1/4000 dilution

    All lanes : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line, treated with 500 ng/ml Trichostatin A for 4 hours. Whole cell lysate.

    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: 14,15 kDa
    Observed band size: 14,15 kDa why is the actual band size different from the predicted?



    This data was developed using ab177863, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 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. Negative 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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse colon tissue is observed. Counter stained with Hematoxylin. Negative 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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 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. Negative 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.
  • Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
    Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab250006? Please let us know so that we can cite the reference in this datasheet.

ab250006 has not yet been referenced specifically in any publications.

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