PARP/Apoptosis Colorimetric Assay Kit

For the measurment of PARP-1 activity in cell extracts
Catalog # Availability Size / Price Qty
4684-096-K

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Product Details
Citations (8)
FAQs
Reviews

PARP/Apoptosis Colorimetric Assay Kit Summary

Ideal for measuring the in vitro activity of PARP-1 in cell extracts before and during apoptosis, screening of candidate PARP-1 inhibitors and determination of IC50 values.
 

Key Benefits

• 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.

Product Specifications

  • Measure PARP-1 activity in cell extracts before and after apoptosis.
  • For the screening of candidate PARP-1 inhibitors and determination of IC50 values.

Kit Contents

• 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
• TACS-Sapphire

Specifications

Shipping Conditions
The components for this kit may require different storage/shipping temperatures and may arrive in separate packaging. Upon receipt, store products immediately at the temperature recommended on the product labels.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Species
Multi-species

Limitations

For research use only. Not for diagnostic use.

Product Datasheets

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Citations 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.

8 Citations: Showing 1 - 8
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  1. Cell stress response impairs de novo NAD+ biosynthesis in the kidney
    Authors: Y Bignon, A Rinaldi, Z Nadour, V Poindessou, I Nemazanyy, O Lenoir, B Fohlen, P Weill-Rayn, A Hertig, A Karras, P Galichon, M Naesens, D Anglicheau, PE Cippà, N Pallet
    JCI Insight, 2022;7(1):.  2022
  2. Surface-directed engineering of tissue anisotropy in microphysiological models of musculoskeletal tissue
    Authors: MJ Mondrinos, F Alisafaei, AY Yi, H Ahmadzadeh, I Lee, C Blundell, J Seo, M Osborn, TJ Jeon, SM Kim, VB Shenoy, D Huh
    Science Advances, 2021;7(11):.  2021
  3. Hydroquinone Induces NLRP3-Independent IL-18 Release from ARPE-19 Cells
    Authors: N Bhattarai, E Korhonen, Y Mysore, K Kaarnirant, A Kauppinen
    Cells, 2021;10(6):.  2021
  4. Animal Evidence for Synergistic Induction of Hepatic Injury by Dietary Fat and Alcohol Consumption and Its Potential Mechanisms
    Authors: HG Kim, JH Wang, HS Kim, JS Lee, HJ Im, SB Lee, DS Lee, GM Hur, CG Son
    Journal of personalized medicine, 2021;11(4):.  2021
  5. Muscle NAD+ depletion and Serpina3n as molecular determinants of murine cancer cachexia - the effects of blocking myostatin and activins
    Authors: JJ Hulmi, F Penna, N Pöllänen, TA Nissinen, J Hentilä, L Euro, JH Lautaoja, R Ballarò, R Soliymani, M Baumann, O Ritvos, E Pirinen, M Lalowski
    Mol Metab, 2020;0(0):101046.  2020
  6. Molecular signatures of BRCAness analysis identifies PARP inhibitor Niraparib as a novel targeted therapeutic strategy for soft tissue Sarcomas
    Authors: H Li, J Tu, Z Zhao, L Chen, Y Qu, H Li, H Yao, X Wang, DF Lee, J Shen, L Wen, G Huang, X Xie
    Theranostics, 2020;10(21):9477-9494.  2020
  7. Poly(ADP-Ribose) Links the DNA Damage Response and Biomineralization
    Authors: KH Müller, R Hayward, R Rajan, M Whitehead, AM Cobb, S Ahmad, M Sun, I Goldberga, R Li, U Bashtanova, AM Puszkarska, DG Reid, RA Brooks, JN Skepper, J Bordoloi, WY Chow, H Oschkinat, A Groombridg, OA Scherman, JA Harrison, A Verhulst, PC D'Haese, E Neven, LM Needham, SF Lee, CM Shanahan, MJ Duer
    Cell Rep, 2019;27(11):3124-3138.e13.  2019
  8. 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

FAQs

  1. 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.

  2. Does the PARP/Apoptosis Colorimetric Assay kit detect mono-ribosylation or poly-ribosylation?

    • This kit recognizes PAR polymers 2 to 50 units long. 

  3. Can the PARP/Apoptosis Colorimetric Assay Kit be used for human samples?

    • Yes, the kit is designed for multiple species including human samples.

  4. 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. 

  5. 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.

  6. Does the standard curve always need to be included when the PARP/Apoptosis Colorimetric Assay Kit is tested?

    • R&D Systems strongly recommends including a standard curve in every assay. The PARP Standard Curve verifies the assay is working properly and allows the user to express the level of PARP activity in their extract as PARP units per amount of protein or PARP units per cell number.

  7. What does the PARP/Apoptosis Colorimetric Assay kit measure?

    • This kit is designed to measure PARP activity before and after apoptosis. It can be used for the screening of candidate PARP-1 inhibitors and determination of IC50 values.

  8. Is it possible to purchase a smaller version of the PARP/Apoptosis Colorimetric Assay Kit?

    • R&D Systems does not offer a smaller version of this kit. However, plates are provided in stripwell format, so the user is able to test fewer wells and save the remaining plate strips for later use.

  9. Is it possible to test partial plates when using the PARP/Apoptosis Colorimetric Assay Kit?

    • The histone-coated plates are provided in stripwell format. To run partial plates, remove excess microplate strips, seal in the provided pouch, and store at 2-8 °C with a dessicant.

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