KEAP1:Nrf2 Assay Service
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Target
KEAP1:Nrf2
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Description
Screening and/or profiling compounds that inhibit the interaction between Keap1 and Nrf2 in a biochemical assay.
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Synonyms
Kelch-like ECH-associated protein 1, Cytosolic inhibitor of Nrf2, INRF2, Kelch-like protein 19, KIAA0132, KLHL19
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Example Data
*Example only, final data may vary.
Assay Details
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Assay Format
Fluorescence Polarization
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Reference Compounds and IC50
KI696, 0.93 μM
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Assay Principle
The Keap1:Nrf2 Inhibitor Screening Assay is designed for identification of inhibitors of Keap1:Nrf2 binding using fluorescence polarization (FP), a powerful tool for studying molecular interactions by monitoring the change in rotational mobility of the fluorescently-labeled molecules upon their binding to a partner. To determine the effect of the inhibitor on the formation of Keap1:Nrf2 complexes, the Keap1 protein and the fluorescent Nrf2 peptide are incubated with or without the test inhibitor for 30 minutes. Changes in rotational mobility of the Nrf2 peptide are measured using a plate reader capable of measuring fluorescence polarization.
Target Details
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Protein Family
Ubiquitination
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UniProt
Q14145
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Background
Nuclear factor erythroid 2-related factor (Nrf2) is a transcription factor that coordinates antioxidant and anti-inflammatory responses by increasing the expression of antioxidant proteins through binding to the ARE (antioxidant response element) region of their promoters. Under basal conditions, Nrf2 is retained in the cytosol by cytoskeletal Keap1, a substrate recognition subunit of a E3 ligase complex which targets Nrf2 for degradation. Exposure to oxidative stress causes the release of Nrf2 from Keap1. Nrf2 translocates to the nucleus, where it binds to AREs and induce the expression of antioxidant and phase II proteins protecting the cell from oxidative damage. Both Nrf2 and Keap1 are attractive therapeutic targets.
Delivery
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Estimated Turnaround
Two to three weeks following delivery of compounds
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Results
Extensive report with raw and analyzed data, graphs, and detailed protocols. Includes positive control for inhibition.
