KEAP1-Nrf2 Inhibitor Screening Assay Kit

Catalog #

72020

$960 *

Size: 96 reactions

Qty

Bulk/Custom

Add to Favorites

*US Pricing only. For international pricing, please contact your local distributor.

●

Purchase

●

Description

The Keap1:Nrf2 Inhibitor Screening Assay Kit 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.

Figure 1. Illustration of the Keap1:Nrf2 Inhibitor Screening Assay Kit.

This product has been cited 12 times.

●

Synonyms

Kelch-like ECH-associated protein 1, Cytosolic inhibitor of Nrf2, INRF2, Kelch-like protein 19, KIAA0132, KLHL19

●

Product Data Gallery

Left: Binding of Keap1 to Nrf2. Right: inhibition of Keap1:Nrf2 binding by Nrf2 inhibitor

Product Info

Storage and Usage

Citations12

Assay Kit Format

Fluorescence Polarization

Supplied As

The 96-well format assay kit contains sufficient amounts of purified Keap1 protein, fluorochrome-conjugated Nrf2 peptide containing the ETGE motif, and assay buffer for 100 binding reactions.

Materials Required But Not Supplied

Adjustable micropipettor and sterile tips
Plate reader capable of fluorescence polarization measurements, λex = 475-495 nm, λem = 518-538 nm.

Format

Catalog #	Name	Amount	Storage
70040	Keap1, Human Recombinant*	30 µg	-80°C	Avoid freeze/ thaw cycles
79112	FAM-Nrf2 peptide (10 µM)	10 µl	-80°C
79113	Keap1-Nrf2 Assay Buffer	10 ml	-20°C
	BSA, 10 mg/ml	100 µl	-80°C
79685	96-well black plate	1	Room Temp

*The concentration of the protein is lot-specific and will be indicated on the tube

UniProt #

Q14145

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.

References

Inoyama, D., et al. 2012. J. Biomol. Screening 17(4): 435-447

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Applications

Great for studying enzyme kinetics and screening small molecule inhibitors for drug discovery and high-throughput applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]

Warnings

This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications.  

Avoid freeze/thaw cycles. Protect Nrf2-FITC from light.

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Ubiquitination

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

KEAP1-Nrf2 Inhibitor Screening Assay Kit

PRODUCTS

SERVICES

RESOURCES

COMPANY