Sample experiment using ab131382 on HeLa cells following drug treatment: H2A.X (pSer139) 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.
Specificity of H2A.X (pSer139) 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 (pSer139) in the untreated cells.
All lanes : H2A.X (pSer139) Human In-Cell ELISA Kit (IR) (ab131382)Lane 1 : Untreated cellsLane 2 : Camptothecin treated cellsLane 3 : UV exposed cells
Antibody specificity demonstrated by Western Blot Analysis: H2A.X (pSer139) is phospho-specific. Jurkat cells were stimulated with UV light exposure to induce H2A.X phospho Ser139 and then the UV treated lysate was treated with lambda protein phosphatase. Top panel: H2A.X (pSer139) is induced by UV treatment and the western blot band is sensitive to phosphatase treatment. Lower panel: In contrast, total H2A.X (ab124781) levels are not sensitive to phosphatase treatment.