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)
• Precision

  Intra-assay

  Sample	n	Mean	SD	CV%
  HeLa Cells				< 10%
• Tests
  1 x 96 well plate
• Sample type
  Adherent cells, Suspension cells
• Assay typeCell-based
• 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).

• Tested applicationsIn-Cell ELISA more details

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	1 x 96 tests
  1000X IRDye-labeled Secondary Antibodies	1 x 24µl
  100X Anti-cleaved-PARP Primary Antibody (Mouse Monoclonal)	1 x 120µl
  100X Anti-H2A.X (pSer139) Rabbit Primary Antibody	1 x 120µl
  100X Triton X-100	1 x 0.5ml
  10X Blocking Buffer	1 x 10ml
  10X Phosphate Buffered Saline	1 x 100ml
  400X Tween-20	1 x 2ml
  Janus Green Stain	1 x 17ml

• Research Areas
  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Apoptosis

  • Kits/ Lysates/ Other
  • Kits
  • ELISA Kits
  • In Cell ELISA Kits
  • Phosphoprotein ICE kits

  • Epigenetics and Nuclear Signaling
  • Histones
  • H2A
  • Phosphorylated

  • Epigenetics and Nuclear Signaling
  • Histones
  • H2A
  • Variants

  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • DNA Damage & Repair
  • Base Excision Repair

  • Epigenetics and Nuclear Signaling
  • Chromatin Modifying Enzymes
  • ADP-ribosylation

  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Apoptosis
  • Nuclear

  • Cell Biology
  • Apoptosis
  • Nucleus
  • PARP

Applications

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

• 

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

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

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

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

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

Protocol Booklet