NRF2 Pathway

NRF2 [Nuclear factor (erythroid-derived 2)-like 2] and it's downstream target genes are the primary cellular defense against the cytotoxic effects of oxidative stress.  Cell damage due to free radicals from oxidative stress contributes to diseases such as arteriosclerosis, stroke, asthma, cardiac infarctions, Alzheimer's, Parkinson's & cancer making the NRF2 pathway a high profile therapeutic target.

Eurofins DiscoverX offers a target-specific, functional cellular assay for NRF2 translocation. This simple mix-and-read assay provides a non-transcriptional response based on NRF2 translocation to the nucleus upon activation.

 

Features of PathHunter Keap1-NRF2 assay
 

  • Functional, cell-based assay

  • Non-transcriptional readout to minimize false positives
  • Simple protocol, rapid results
  • HTS-compatible assay
  • Available  as cell lines or a convenient thaw-n-use format
 

Assay List

Cell Lines

Target Description Cat. No.
Keap1-NRF2 PathHunter® U2OS Keap1-NRF2 Nuclear Translocation Cell Line 93-0821C3

Assay Ready Kits
Target Description Cat. No.
Keap1-NRF2 PathHunter® eXpress Keap1-NRF2 Nuclear Translocation Assay 93-0821E3CP0L
Keap1-NRF2 PathHunter® eXpress Keap1-NRF2 Nuclear Translocation Assay 93-0821E3CP0M
Keap1-NRF2 PathHunter® eXpress Keap1-NRF2 Nuclear Translocation Assay 93-0821E3CP0S

 


Assay Principle

The PathHunter NRF2 assay is based on the Enzyme Fragment Complementation (EFC) technology.  A small inactive fragment of β-gal (PK tag) was fused to the NRF2 protein in a recombinant cell line. The other inactive fragment of β-gal, termed EA for ‘enzyme acceptor’, is anchored in the nucleus. Activation of the Keap1-NRF2 protein complex results in NRF2 stabilization and translocation to the nucleus, resulting in PK-EA complementation and formation of a functional enzyme. Enzyme activity is then quantitatively detected using a chemiluminescent substrate in the PathHunter® Detection Kit.

How it Works

PathHunter KEAP1-NRF2 Translocation Assay