MitoSciences (MS1098)

H2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit (IR) (ab131383)

Overview

  • Product nameH2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit (IR)
  • Detection methodIR
  • Tests
    1 x 96 well plate
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeCell-based (qualitative)
  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    The H2A.X (pS139) and cleaved-PARP Human In-Cell ELISA Kit (IR) (ab131383) is designed to study the induction of DNA damage and/or apoptosis in response to various stimuli. Monoclonal antibodies specific to phospho-H2A.X (S139) and cleaved-PARP are used in the high-throughput duplexing plate-based assay. H2A.X is a histone H2A family member that is phosphorylated and recruited to sites of double-strand DNA breaks. Poly [ADP-ribose] polymerase 1 (PARP) is a DNA repair enzyme that is cleaved by activated caspases. Combined, these antibodies provide biomarkers of dsDNA breaks (pH2A.X) and apoptosis (cleaved-PARP).

     

    Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately.

  • Tested applicationsIn-Cell ELISA more details
  • PlatformMicroplate

Properties

Applications

Our Abpromise guarantee covers the use of ab131383 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
In-Cell ELISA Use at an assay dependent concentration.

H2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit (IR) images

  • Sample experiment using ab131383 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 data (presented as mean +/- SD) were analyzed as described in Data Analysis section. The dashed grey line indicates the vehicle control signal.
  • Sample experiment using ab131383 on HeLa cells following drug treatment: cleaved-PARP readout. HeLa cells were treated for 4 hours with dose titrations of Camptothecin, Etoposide and Staurosporin. The data (presented as mean +/- SD) were analyzed as described in Data Analysis section. The dashed grey line indicates the vehicle control signal.
  • Specificity of H2A.X (pSer139) and cleaved-PARP antibodies demonstrated by immunocytochemistry. Primary antibodies used in this assay kit were 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) and cleaved-PARP signal in the untreated cells.
  • All lanes : H2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit (IR) (ab131383)

    Lane 1 : Untreated cells
    Lane 2 : Camptothecin treated cells
    Lane 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 (pSer139) 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.

Protocols

References for H2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit (IR) (ab131383)

ab131383 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"