Revvity, 64PARPEG, HTRF Human and Mouse PARP cleaved-Asp214 Detection Kit, 500 Assay Points

Overview
The cleaved PARP (Asp214) kit enables the specific and quantitative detection of endogenous levels of the human, mouse, and monkey large fragment (89-kDa) of PARP-1. With its central position downstream from caspases 3 and 7, both activated by extrinsic and intrinsic apoptosis pathways, cleaved PARP-1 represents a pertinent marker of cells undergoing apoptosis. Deregulated apoptosis plays a role in various diseases involving insufficient cell death, such as cancer, autoimmunity, and persistent infections, whereas excessive apoptosis can contribute to neuro-degeneration and ischaemia.
How it works
Assay principle

 
The cleaved-PARP assay is a sandwich immunoassay using two specific anti-PARP-1 p85 fragment monoclonal antibodies, one labelled with Eu3+ Cryptate (donor) and the other with d2 (acceptor). Donor-acceptor proximity leads to a fluorescent TR-FRET signal proportional to the number of cells undergoing apoptosis. The protocol is optimized for a 384-well plate format, but can easily be further miniaturized or upscaled. After cell lysis, the cleaved PARP-1 p85 fragment can be semi-quantitatively detected using the HTRF cleaved-PARP-Asp214 assay kit reagents.

Assay protocol
Two-plate assay protocol For added flexibility, the assay can be run under a two-plate assay protocol, where cells are plated and treated in a 96-well culture plate. For detection, lysates are subsequently transferred to a 384-well sv assay plate where the detection reagents are added. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step. This also enables monitoring of the cells' viability and confluence in an appropriate cell culture plate. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Cardiovascular Infectious Diseases Metabolism/Diabetes NASH/Fibrosis Neuroscience Rare Diseases
Unit Size-500 assay points