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.

Can samples containing protease inhibitors be used?

Protease inhibitor PMSF has been validated for use in this kit. Other protease inhibitors may be suitable and would require end user evaluation.

PARP/Apoptosis Colorimetric Assay Kit

Name: PARP/Apoptosis Colorimetric Assay Kit
Brand: R&D Systems
SKU: 4684-096-K

R&D Systems, part of Bio-Techne | Catalog # 4684-096-K

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Citations (13)

Product Datasheet / COA

For the measurment of PARP-1 activity in cell extracts

Key Product Details

Features:

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

Technical Data
Citations / Reviews / FAQs

Summary for PARP/Apoptosis Colorimetric Assay Kit

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 for PARP/Apoptosis Colorimetric Assay Kit

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

Product Specifications for PARP/Apoptosis Colorimetric Assay Kit

Species

Multi-species

Preparation & Storage

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.

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PARP/Apoptosis Colorimetric Assay Kit

R&D Systems, part of Bio-Techne | Catalog # 4684-096-K

Key Product Details

Summary for PARP/Apoptosis Colorimetric Assay Kit

Why Use the PARP/Apoptosis Colorimetric Assay Kit?

Product Specifications

Kit Contents for PARP/Apoptosis Colorimetric Assay Kit

Product Specifications for PARP/Apoptosis Colorimetric Assay Kit

Preparation & Storage

Customer Reviews for PARP/Apoptosis Colorimetric Assay Kit

FAQs for PARP/Apoptosis Colorimetric Assay Kit

Product Documents for PARP/Apoptosis Colorimetric Assay Kit

Product Datasheets

COA

Certificate of Analysis

CONTACT INFORMATION

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