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Product name
PARP-1 (cleaved) In-Cell ELISA Kit (IR)
Specificity
The PARP-1 (cleaved) In-Cell ELISA Kit (IR) is a highly specific and high-throughput assay for measuring the larger (89 kDa) fragment of poly (ADP-ribose) polymerase 1 (PARP-1) generated by caspase cleavage between Asp214 and Gly215 of human PARP-1. The kit does not recognize full-length PARP-1 or the 24 kDa DNA binding domain fragment.
Tests
2 x 96 well plate
Sample type
Adherent cells, Suspension cells
Assay type
Direct
Cross reactivity
Reacts with
Human
Product overview
In-Cell ELISA Kits use quantitative immunocytochemistry to measure protein levels or post-translational modifications in cultured cells. Cells are fixed in a 96- or 384-well plate and targets of interest are detected with highly specific, well-characterized monoclonal antibodies, and levels are quantified with IRDye®-labeled Secondary Antibodies. IR imaging and quantitation is performed using a LI-COR® Odyssey® or Aerius® system.
In-Cell ELISA (ICE) technology is used to perform quantitative immunocytochemistry of cultured cells with a near-infrared fluorescent dye-labeled detector antibody. The technique generates quantitative data with specificity similar to Western blotting, but with much greater quantitative precision and higher throughput due to the greater dynamic range and linearity of direct fluorescence detection and the ability to run 96 samples in parallel. This method rapidly fixes the cells in situ, stabilizing the in vivo levels of proteins, and thus essentially eliminates changes during sample handling, such as preparation of protein extracts. Finally, the cleaved PARP-1 signal can be normalized to cell amount, measured by the provided Janus Green whole cell stain1, to further increase the assay precision.
The assay is designed for use with cultured adherent and suspension cells in a 96-well microplate format. The adherent cells undergoing apoptosis readily detach from a culture plate. The cell detachment often leads to their loss and thus underestimating the proportion of apoptotic cells. This assay was developed to eliminate the loss of apoptotic cells. In addition, this assay is also applicable for suspension cells. Materials and separate protocol sections are provided to achieve efficient attachment of apoptotic adherent cells as well as suspension cells.
LI-COR®, Odyssey®, Aerius® and IRDye® are registered trademarks of LI-COR Biosciences Inc.
Tested applications
In-Cell ELISAmore details
Properties
Storage instructions
Store at 4°C for upto 6 months. Refer to the protocol for further storage instructions.
| Components | 2 x 96 tests |
|---|---|
| 1000X IRDye800-labeled Secondary Antibody | 1 x 24µl |
| 100X Triton X-100 | 1 x 0.5ml |
| 10X Blocking Solution | 1 x 15ml |
| 10X Phosphate Buffered Saline | 1 x 100ml |
| 1X Janus Green Stain | 1 x 11ml |
| 400X Tween-20 | 1 x 2ml |
| 500X Primary Antibody | 1 x 42µl |
| 96-well Assay Plate | 2 units |
| Plate Seals | 2 units |
Research areas
Metabolism >> Pathways and Processes >> Metabolism processes >> Apoptosis
Kits/ Lysates/ Other >> Kits >> Cell Damage Kits >> DNA damage and cell cycle arrest
Cell Biology >> Apoptosis >> Mitochondrial
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> Base Excision Repair
Epigenetics and Nuclear Signaling >> Chromatin Modifying Enzymes >> ADP-ribosylation
Applications
Show applications key
Our Abpromise guarantee covers the use of ab110215 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
In-Cell ELISA
In-Cell ELISAIn-Cell ELISA
PARP-1 (cleaved) In-Cell ELISA Kit (IR) images:
ELISA - PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
Figure 1. Cell number and treatment time-dependence of PARP-1 cleavage in adherent cells induced to undergo apoptosis.
ELISA - PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
Figure 2. Determination of Staurosporine EC50 of PARP-1 cleavage in HeLa cells. HeLa cells were seeded at 50,000 cells per well, allowed to attach overnight and treated with Staurosporine for 4 hours. Cells were fixed and the plate was analyzed by ICE to measure the cleaved PARP-1 using MSA43. All steps were performed as described in MSA43 Protocol. Mean and standard error of the mean (n=2) is shown. The EC50 of Staurosporine is 0.072 µM (R2=0.954).
ELISA - PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
Figure 3. Cell-line dependent induction of PARP-1 cleavage.
Immunocytochemistry/ Immunofluorescence - PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
Figure 4. PARP-1 (cleaved) antibody specificity demonstrated by immunocytochemistry. Immunocytochemical analysis of HeLa cells either untreated or treated with 1 µM Staurosporine (STS) for 4 hours to induce apoptosis. The same PARP-1 (cleaved) antibody as in the MSA43 was used in this analysis (in green). Cells well co-stained with DNA stain DAPI (in red). Note that the PARP-1 (cleaved) antibody specifically labels nucleus of only STS-treated cells
Western blot - PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
Figure 5. PARP-1 (cleaved) antibody specificity demonstrated by Western Blot. Western blot analysis of HeLa cells either untreated (CON) or treated with 1 µM Staurosporine (STS) for 4 hours to induce apoptosis. The same PARP-1 (cleaved) antibody as in the MSA43 was used in this analysis. Note that the antibody recognize the 89 kDa cleaved PARP-1 and it does not recognize the 113 kDa full length PARP-1
Protocols
References for PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215)
ab110215 has not yet been referenced specifically in any publications.
Publishing research using ab110215? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"