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.
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.
PathHunter KEAP1-NRF2 Translocation Assay
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.
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
View Categories: Pathway Assays
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