Human Phospho-Histone H2AX (S139) Antibody Summary
Accession # P16104
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
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 (Catalog # 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 PBMC by Flow Cytometry. Human PBMC untreated (open histogram) or treated with 1 µM Camptothecin (Catalog # 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 (Catalog # F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with 90% methanol. View our protocol for Staining Intracellular Molecules.
Preparation and Storage
- 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.
Background: Histone H2AX
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.
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PC3 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