Nuclear Factor (erythroid-derived 2)-like 2 (Nrf2)
Nrf2 is a ubiquitously expressed, basic leucine zipper transcription factor. It regulates the expression of a variety of genes encoding proteins that play critical roles in cyto-protection, as well as the detoxification and clearance of harmful endogenous and xenobiotic substances. In particular, Nrf2 regulates the expression of antioxidant proteins that confer cyto-protection against oxidative damage.
Under normal conditions Nrf2 resides in the cytoplasm bound to Keap1 and Cullin 3, and is the target of ubiquitination and rapid turn-over via proteasomal degradation. However, under conditions of cellular oxidative stress the tight association of Nrf2 with Keap1 and Cullin 3 is broken, effectively disrupting the otherwise efficient process of Nrf2 degradation. Once non-ubiquitinated Nrf2 accumulates in the cytoplasm it translocates into the nucleus, whereupon it forms hetero-dimers with Maf. In this configuration Nrf2 binds to antioxidant response element (ARE) sequences resident in the promoter regions of some genes, initiating transcription complex formation and gene expression.
INDIGO’s Human Nrf2 Reporter Cells include the luciferase reporter gene functionally linked to a promoter containing tandem antioxidant response elements (AREs). Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in Nrf2 activity. The principle application of this reporter assay system is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human Nrf2.
Kits are offered in different assay formats to accommodate researchers’ needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
INDIGO’s Human Nrf2 Assay kit is an all-inclusive, single use assay system. In addition to Nrf2 Reporter Cells, kits provide optimized media for use during cell culture and in diluting user’s test samples, a reference activator, Luciferase Detection Reagent, and a cell culture-ready assay plate.
Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay wells. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments prior to assay setup. Reporter Cells are single use reagents. They can not be refrozen or maintained in extended culture.
| Product Family | Product Number | Product Description |
| IB1000 Nrf2 | IB10001-32 | Nrf2 Reporter Assay System, 3 x 32 assays in 96-well format |
| IB10001 | Nrf2 Reporter Assay System, 1 x 96-well format assays | |
| IB10002 | Nrf2 Reporter Assay System, 1 x 384-well format assays |
Service Assays: Human
The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. Service assays include a positive control reference compound and ‘vehicle’ control for every experiment.A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online Request a Quote form or contact an INDIGO Customer Service Representative to discuss your desired study parameters. To initiate a Service Study, download and complete all fields of the Excel worksheet “Service Work Order" then submit the electronic file to INDIGO Customer Service.
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