PARP/Apoptosis Colorimetric Assay Kit
PARP/Apoptosis Colorimetric Assay Kit SummaryIdeal 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.
• 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 diagnostic use.
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.
Citations: Showing 1 - 10
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Nuclear DJ-1 Regulates DNA Damage Repair via the Regulation of PARP1 Activity
Authors: Wang, ZX;Liu, Y;Li, YL;Wei, Q;Lin, RR;Kang, R;Ruan, Y;Lin, ZH;Xue, NJ;Zhang, BR;Pu, JL;
International journal of molecular sciences 2023-05-12
NAD+ repletion with niacin counteracts cancer cachexia
Authors: M Beltrà, N Pöllänen, C Fornelli, K Tonttila, MY Hsu, S Zampieri, L Moletta, S Corrà, PE Porporato, R Kivelä, C Viscomi, M Sandri, JJ Hulmi, R Sartori, E Pirinen, F Penna
Nature Communications, 2023-04-03;14(1):1849. 2023-04-03
p-Hydroxybenzyl Alcohol Antagonized the ROS-Dependent JNK/Jun/Caspase-3 Pathway to Produce Neuroprotection in a Cellular Model of Parkinson's Disease
Authors: MC Lai, WY Liu, SS Liou, IM Liu
Nutrients, 2022-11-24;14(23):. 2022-11-24
Serine metabolism remodeling after platinum-based chemotherapy identifies vulnerabilities in a subgroup of resistant ovarian cancers
Authors: T Van Nyen, M Planque, L van Wagens, JAG Duarte, EA Zaal, A Talebi, M Rossi, PR Körner, L Rizzotto, S Moens, W De Wispela, REM Baiden-Ami, GS Sonke, HM Horlings, G Eelen, E Berardi, JV Swinnen, CR Berkers, P Carmeliet, D Lambrechts, B Davidson, R Agami, SM Fendt, D Annibali, F Amant
Nature Communications, 2022-08-05;13(1):4578. 2022-08-05
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, 2022-01-11;7(1):. 2022-01-11
Hydroquinone Induces NLRP3-Independent IL-18 Release from ARPE-19 Cells
Authors: N Bhattarai, E Korhonen, Y Mysore, K Kaarnirant, A Kauppinen
Cells, 2021-06-06;10(6):. 2021-06-06
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, 2021-04-08;11(4):. 2021-04-08
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, 2021-03-12;7(11):. 2021-03-12
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, 2020-07-25;10(21):9477-9494. 2020-07-25
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, 2020-06-26;0(0):101046. 2020-06-26
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, 2019-06-11;27(11):3124-3138.e13. 2019-06-11
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, 2012-09-19;2(12):1134-49. 2012-09-19
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.
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.
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.
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.
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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