Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) is Cell-based (qualitative) ELISA for the measurement of Human H2A.X (phospho S139) (IR) production in Human in Suspension cells, Adherent cells samples.
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- Current issues in molecular biology 45:7915-79322023DNA Double-Strand Break Repair Inhibitors: YU238259, A12B4C3 and DDRI-18 Overcome the Cisplatin Resistance in Human Ovarian Cancer Cells, but Not under Hypoxia Conditions.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAnna Macieja,Izabela Gulbas,Tomasz PopławskiPubMed 37886943
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- Molecular cancer therapeutics 13:2817-262014Antitumor effects of a novel small molecule targeting PCNA chromatin association in prostate cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKelsey L Dillehay et. al.PubMed 25253786
Key facts
- Detection method
- IR
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based (qualitative)
- Reactive species
- Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
What's included?
Recommended products
Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) is Cell-based (qualitative) ELISA for the measurement of Human H2A.X (phospho S139) (IR) production in Human in Suspension cells, Adherent cells samples.
Key facts
- Detection method
- IR
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based (qualitative)
- Reactive species
- Human
- Assay Platform
- Microplate
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample HeLa Extract | n 0 | mean - | SD - | C.V. < 10 |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
The H2A.X (phospho S139) Human In-Cell ELISA Kit (IR) (ab131382) is designed to study the induction of DNA damage in response to various stimuli. A rabbit monoclonal antibody specific to H2A.X phospho S139 is used in this high-throughput duplexing plate-based assay. H2A.X (phospho S139) is a reliable readout for double-stranded DNA breaks.
Plates are available in our ICE (In-Cell ELISA) Support Pack (In-Cell ELISA (ICE) Support Pack ab111542) which can be bought seperately.
In-Cell ELISA (ICE) technology is used to perform quantitative immunocytochemistry of cultured cells with a near-infrared fluorescent dye-labeled detector antibody. The technique generates quantitative data with specificity similar to western blotting, but with much greater quantitative precision and higher throughput due to the greater dynamic range and linearity of direct fluorescence detection and the ability to run up to 96 samples in parallel.
This method rapidly fixes the cells in situ, stabilizing the in vivo levels of proteins and their posttranslational modifications, and thus essentially eliminates changes during sample handling, such as preparation of protein extracts. Finally, the H2A.X (phospho S139) signal can be normalized to cell amount, measured by the provided Janus Green whole-cell stain, to further increase the assay precision.
This product is designed for LI-COR® Odyssey® or Aerius® infrared imaging systems.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The protein target H2A.X pSer139 also known as phospho-histone H2A.X and H2AX is a modified form of histone H2A.X which occurs when serine 139 becomes phosphorylated. This histone variant with a mass of approximately 15 kDa plays an important role in the DNA damage response. You can find H2A.X pSer139 in the chromatin of eukaryotic cells. It is especially abundant in tissues actively dividing like bone marrow and lymphoid tissues where it responds to DNA double-strand breaks by undertaking a structural change that signals the recruitment of repair proteins to the site of damage.
- Biological function summary
H2A.X pSer139 serves as an indicator of DNA repair processes and contributes to maintaining genomic stability. When DNA damage occurs H2A.X rapidly phosphorylates and forms part of a complex including other repair proteins like MDC1 and 53BP1 marking regions for DNA repair machinery. By localizing with damaged DNA H2A.X pSer139 helps prevent genome instability by coordinating repair responses such as homologous recombination and non-homologous end joining.
- Pathways
H2A.X pSer139 plays a significant role in the DNA damage response (DDR) and cell cycle checkpoint pathways. Within the ATM/ATR signaling pathway phosphorylation of H2A.X is an early step triggering cellular checkpoint controls and DNA repair. Proteins like ATM a kinase activated by DNA damage interact with H2A.X pSer139 phosphorylating it to initiate recruitment of repair complexes. Furthermore H2A.X pSer139 links with CHK2 a protein kinase in the DDR pathway helping regulate the cell cycle in response to DNA damage.
- Associated diseases and disorders
Alterations in H2A.X pSer139 function associate with cancer and neurodegenerative disorders. In many cancers defective DNA damage response often involving aberrations in H2A.X phosphorylation leads to genomic instability and tumor progression. In neurodegenerative diseases H2A.X pSer139 often coexists with proteins such as tau where DNA repair deficiencies contribute to neuronal damage. Understanding the regulation of H2A.X pSer139 offers insights into diseases and potential therapeutic targets.
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4 product images
Western blot - Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) (ab131382)
Antibody specificity demonstrated by Western Blot Analysis: H2A.X (phospho S139) is phospho-specific. Jurkat cells were stimulated with UV light exposure to induce H2A.X (phospho S139) and then the UV treated lysate was treated with lambda protein phosphatase. Top panel: H2A.X (phospho S139) is induced by UV treatment and the western blot band is sensitive to phosphatase treatment. Lower panel: In contrast, total H2A.X (Anti-Histone H2A.X antibody [EPR895] - Nuclear Marker ab124781) levels are not sensitive to phosphatase treatment.
Western blot - Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) (ab131382)
Antibody specificity demonstrated by Western Blot Analysis. Whole cell lysates from Jurkat cells were analyzed by western blot with the H2A.X (phospho S139) antibody used in this assay kit.
Immunocytochemistry/ Immunofluorescence - Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) (ab131382)
Specificity of H2A.X (phospho S139) antibodies demonstrated by immunocytochemistry. The primary antibody used in this assay kit was validated by staining HeLa cells treated with 10 μM Camptothecin or vehicle for 4 hours and imaged by fluorescent microscopy. Note the absence of H2A.X (phospho S139) in the untreated cells.
In-Cell ELISA - Human H2A.X (phospho S139) In-Cell ELISA Kit (IR) (ab131382)
Sample experiment using ab131382 on HeLa cells following drug treatment: H2A.X (phospho S139) readout. HeLa cells were treated for 4 hours with dose titrations of Camptothecin, Etoposide and Staurosporin. The dashed grey line indicates the vehicle control signal.
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