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Rabbit Recombinant Monoclonal H2A.X antibody. Nuclear Membrane marker. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 31 publications.


Images

Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (AB124781), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (AB124781), expandable thumbnail
  • Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (AB124781), expandable thumbnail
  • Immunoprecipitation - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (AB124781), expandable thumbnail
  • Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (AB124781), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Rat
Expected
Expected
Tested
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/500 - 1/1000

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

5 µg/mL

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000 - 1/10000

Notes

-

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

-

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

ab124781 does not work on PFA fixed cells but does work on methanol fixed cells.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

7 products for Alternative Product

Target data

Function

Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. 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. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.

Alternative names

Histone H2AX, H2a/x, Histone H2A.X, H2AX, H2AFX

Recommended products

Rabbit Recombinant Monoclonal H2A.X antibody. Nuclear Membrane marker. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 31 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR895

Purification technique

Affinity purification Protein A

Specificity

This antibody does not work in ICC/IF on PFA fixed cells but does work on methanol fixed cells. The immunogen used for this product shares full homology with other H2A proteins, including H2A1, H2A1B, H2A2A and H2AZ. Cross-reactivity with these proteins has not been confirmed experimentally.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

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 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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781) at 1/3000 dilution

    Lane 1: Raji cell lysate at 20 µg

    Lane 2: HEK293 cell lysate at 20 µg

    Lane 3: Mouse kidney tissue lysate at 20 µg

    Lane 4: Rat kidney tissue lysate at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 15 kDa

    Observed band size: 14 kDa, 22 kDa

  • Flow Cytometry (Intracellular) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Histone H2A.X with purified ab124781 at 1/100 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    ~22kDa band may be the monoubiquitinated form of histone H2A

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781) at 1/2000 dilution

    Lane 1: Raji cell lysate at 20 µg

    Lane 2: HEK293 cell lysate at 20 µg

    Lane 3: Mouse kidney tissue lysate at 20 µg

    Lane 4: Rat kidney tissue lysate at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 15 kDa

    Observed band size: 14 kDa, 22 kDa

  • Immunoprecipitation - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Immunoprecipitation - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Histone H2A.X was immunoprecipitated using 5ug of ab124781 from 200ul of HeLa whole cell extract lysate diluted to 0.5mg/ml in RIPA and 50ul of Protein G magnetic beads. No antibody was added to the control (-).

    The antibody was incubated under agitation with the Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70°C. 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab124781 at 1ug/ml. The Secondary antibody was Mouse monoclonal SB62a Anti-Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697) at 1/10,000 dilution.

    Band: 15kDa; Histone H2A.X

    All lanes: Immunoprecipitation - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa

    Exposure time: 20min

  • Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    All lanes: Western blot - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781) at 1/1000 dilution

    Lane 1: Raji lysate at 10 µg

    Lane 2: Fetal kidney lysate at 10 µg

    Lane 3: Human heart lysate at 10 µg

    Secondary

    All lanes: HRP labelled Goat anti Rabbit IgG at 1/2000 dilution

    Predicted band size: 15 kDa

    Observed band size: 14 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Histone H2A.X with unpurified ab124781 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Histone H2A.X with purified ab124781 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker (ab124781)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H2A.X with purified ab124781 at a dilution of 1/1000. Cells were fixed with either 4% PFA (top) or 100% methanol (bottom) and permeabilized with 0.1% TritonX-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.

    Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 mouse anti-Tubulin (1/1000) followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were used to label tubulin. DAPI was used as the nulear counterstain.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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