Antibodies, Proteins, Kits and Reagents for Life Science

368 544 ₸ Product size

100 µl 368 544 ₸ 10 µl 150 768 ₸

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR895] to Histone H2A.X
Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt
Reacts with: Mouse, Rat, Human

Product name

Anti-Histone H2A.X antibody [EPR895]
See all Histone H2A.X primary antibodies
Description

Rabbit monoclonal [EPR895] to Histone H2A.X
Host species

Rabbit
Specificity

ab124781 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.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human Histone H2A.X aa 1-100. The exact sequence is proprietary.
Positive control
- WB: Raji and HEK293 cell lysates, fetal kidney, human heart, mouse and rat kidney tissue lysates. IHC-P: Human kidney and clear cell carcinoma tissues. ICC/IF: HeLa cells. IP: HeLa whole cell lysate (ab150035).
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR895
Isotype

IgG
Research areas

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

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

Lane 1 : Raji cell lysate
Lane 2 : HEK293 cell lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Rat kidney tissue lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 15 kDa
Observed band size: 14,22 kDa
why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Flow Cytometry - Anti-Histone H2A.X antibody [EPR895] (ab124781)

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] (ab124781)

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

Lane 1 : Raji cell lysate
Lane 2 : HEK293 cell lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Rat kidney tissue lysate

Lysates/proteins at 20 µg per lane.

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 why is the actual band size different from the predicted?
Additional bands at: 22 kDa. We are unsure as to the identity of these extra bands.

~22kDa band may be the monoubiquitinated form of histone H2A
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Histone H2A.X antibody [EPR895] (ab124781)

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

Lane 1 : Raji lysate
Lane 2 : Fetal kidney lysate
Lane 3 : Human heart lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 15 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody [EPR895] (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] (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] (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. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
ab7291 mouse anti-Tubulin (1/1000) followed by ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were used to label tubulin. DAPI was used as the nulear counterstain.
Immunoprecipitation - Anti-Histone H2A.X antibody [EPR895] (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) (ab99697) at 1/10,000 dilution.
Band: 15kDa; Histone H2A.X
Anti-Histone H2A.X antibody [EPR895] (ab124781)

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