PARP/Apoptosis Colorimetric Assay Kit
PARP/Apoptosis Colorimetric Assay Kit Summary
• Colorimetric readout
• 96 well format
• Highly sensitive – detects 0.1 mU PARP ~500 cells
• Dynamic range between 0.1 to 10 mU PARP
• Requires 10-100 ng extract for detection
• Assay Time ~3 hrs
Why Use the PARP/Apoptosis Colorimetric Assay Kit?
This ELISA based assay detects poly (ADP-ribose) deposited by PARP-1 onto immobilized histones in a 96-well format. An anti-PAR monoclonal detecting antibody followed by addition of a goat anti-rabbit IgG-HRP secondary and a colorimetric HRP substrate yields relative absorbance that correlates with PARP-1 activity. During apoptosis PARP-1 is cleaved with a resulting drop in activity. Etoposide is included as a control apoptosis inducer.
- Measure PARP-1 activity in cell extracts before and after apoptosis.
- For the screening of candidate PARP-1 inhibitors and determination of IC50 values.
• 10X Activated DNA
• PARP-HSA, 10 mUnits/µl
• 20 mM NAD
• Anti-PAR monoclonal Antibody
• Goat anti-mouse IgG-HRP
• 10 mM Etoposide
• 20X I-PAR Assay Buffer
• 5X Antibody Diluent
• Histone-Coated Natural Strip Well Plate, I-PAR
For research use only. Not for diagnositic use.
Citation for PARP/Apoptosis Colorimetric Assay Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
Dual roles of PARP-1 promote cancer growth and progression.
Authors: Schiewer M, Goodwin J, Han S, Brenner J, Augello M, Dean J, Liu F, Planck J, Ravindranathan P, Chinnaiyan A, McCue P, Gomella L, Raj G, Dicker A, Brody J, Pascal J, Centenera M, Butler L, Tilley W, Feng F, Knudsen K
Cancer Discov, 0;2(12):1134-49. 0
Can cells be lysed with RIPA buffer for use in the PARP/Apoptosis Colorimetric Assay Kit?
RIPA buffer is not recommended for use in the PARP/Apoptosis Colorimetric Assay. The recommended lysis buffer for this assay is the Cell Extraction Buffer described in the insert.
Does the PARP/Apoptosis Colorimetric Assay kit detect mono-ribosylation or poly-ribosylation?
This kit recognizes PAR polymers 2 to 50 units long.
Can the PARP/Apoptosis Colorimetric Assay Kit be used for human samples?
Yes, the kit is designed for multiple species including human samples.
What concentration of etoposide do you recommend to use in the PARP/Apoptosis Colorimetric Assay?
We recommend using less than 5 uM of etoposide for 72 hours. Optimal concentration and incubation time may vary and should be optimized by each laboratory.
Why is it necessary to use two different wash buffer formulations to perform washing in the PARP/Apoptosis Colorimetric Assay procedure?
The wash buffer containing PBS-Triton results in foaming which is difficult to remove without subsequent PBS washes. Signal intensity and reproducibility are also decreased if PBS-Triton wash is not followed up with a PBS wash.
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