NRF2 Pathway
NRF2 [Nuclear factor (erythroid-derived 2)-like 2] and it's downstream target genes play an important role in cellular anti-oxidant defense, making it a therapeutic target for neurodegenerative diseases and cancer. Variety of environmental factors such as smoking, inflammation, grilled meat, sunlight and oxygen lead to production of free radicals and other highly reactive substances that result in cell damage. Cell damage contributes to diseases such as arteriosclerosis, stroke, asthma, cardiac infarctions, Alzheimer's disease, Parkinson's disease and cancer. DiscoveRx offers novel direct, functional cellular assays for NRF2 translocation. No more assay development required; a simple mix-and-read micro titer plate-based assay without obtaining gene transcription is now available!
Features and Benefits
-
Functional, cell-based assay for Keap-NRF2
-
Simple HTS friendly protocol
-
Non-reporter gene based assays minimizes false positives and toxicity
Applications
-
Screen or profile compounds against Keap-NRF2 pathway
-
Obtain toxicity profile of your compounds
-
Screen for: small molecules, siRNA
Assay Principle
PathHunter® cell lines feature novel in vivo applications of Enzyme Fragment Complementation (EFC) technology in which the β-galactosidase (β-gal) enzyme has been split into two inactive fragments. Addition of quinine compounds initiate the release of NRF-2 from KEAP1. This action results in the stabilization of NRF-2 and subsequent nuclear translocation of NRF-2 which results in gene transcription. DiscoveRx has designed an EFC assay to monitor nuclear translocation of NRF -2. A small 4 kDa complementing fragment of β-gal (i.e., the ProLink™ tag) has been fused recombinantly to the NRF2 protein. The larger portion of β-gal, termed EA for ‘enzyme acceptor’, is anchored in the nucleus. Activation of the Keap1-NRF2 protein results in NRF2 translocation that results in PK-EA complementation and formation of a functional enzyme. Enzyme activity is then quantitatively detected using the chemiluminescent substrate in the PathHunter® Detection Kit.
View Categories: Pathway Assays
NRF2 Pathway Related Products