Histone H2AX is a core histone protein that is phosphorylated at S139 in cells exposed to DNA double-strand break-inducing agents, such as ionizing radiation. The S139 phosphorylated H2AX, termed gamma -H2AX, marks the site of DNA double-strand breaks and serves to recruit cell cycle checkpoint and DNA repair factors to the site of damage.
Human phospho-Histone H2AX (S139) Antibody (2207D)
R&D Systems | Catalog # MAB2288
Recombinant Monoclonal Antibody.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Mouse
Applications
Validated:
Western Blot, Intracellular Staining by Flow Cytometry, CyTOF-ready
Cited:
Immunohistochemistry
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # 2207D
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Product Specifications
Immunogen
Phospho-peptide containing human Histone H2AX S139 site
Accession # P16104
Accession # P16104
Specificity
Detects human Histone H2AX when phosphorylated at S139 in Western blots.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human phospho-Histone H2AX (S139) Antibody (2207D)
Detection of Human Histone H2AX by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 20 mJ/cm2 ultraviolet light (UV) followed by a 30 minute recover. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Monoclonal Antibody (Catalog # MAB2288) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Histone H2AX at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-Histone H2AX (S139) in Camptothecin-treated Jurkat cells by Flow Cytometry.
Naïve Jurkat cells (open histogram) or treated with 1 µM Camptothecin (1100, filled histogram) overnight were stained with Rabbit Anti-Human Phospho-Histone H2AX (S139) Monoclonal Antibody (Catalog # MAB2288), followed by PE-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with 90% methanol. View our protocol for Staining Intracellular Molecules.Applications for Human phospho-Histone H2AX (S139) Antibody (2207D)
Application
Recommended Usage
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
Sample: Camptothecin-treated Jurkat cells, fixed with Flow Cytometry Fixation Buffer (Catalog# FC004) and permeabilized with 90% methanol
Sample: Camptothecin-treated Jurkat cells, fixed with Flow Cytometry Fixation Buffer (Catalog# FC004) and permeabilized with 90% methanol
Western Blot
0.25 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with ultraviolet light (UV)
Sample: HeLa human cervical epithelial carcinoma cell line treated with ultraviolet light (UV)
Reviewed Applications
Read 1 review rated 5 using MAB2288 in the following applications:
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Protein A or G purified from cell culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Histone H2AX
Additional Histone H2AX Products
Product Documents for Human phospho-Histone H2AX (S139) Antibody (2207D)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human phospho-Histone H2AX (S139) Antibody (2207D)
For research use only
Related Research Areas
Citations for Human phospho-Histone H2AX (S139) Antibody (2207D)
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Customer Reviews for Human phospho-Histone H2AX (S139) Antibody (2207D) (1)
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Application: Western BlotSample Tested: PC-3 human prostate cancer cell lineSpecies: HumanVerified Customer | Posted 07/16/2018PC3 cells were treated with UV light for 1h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1mm/ml Human Phospho-Histone H2AX (S139) (MAB2288). A specific band was detected for p-H2AX at approximately 17 kDa. This experiment was conducted under reducing conditions
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Liperfluo
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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