Nuclear Factor (erythroid-derived 2)-like 2 (Nrf2)
| Product Family | Product Number | Product Description | Technical Manual |
| IB1000 Nrf2 |
IB10001-32 | Human Nrf2 Reporter Assay System, 3 x 32 assays in 96-well format | Technical Manual |
| IB10001 | Human Nrf2 Reporter Assay System, 1 x 96-well format assays | Technical Manual | |
| IB10002 | Human Nrf2 Reporter Assay System, 1 x 384-well format assays |
Nuclear Factor (erythroid-derived 2)-like 2 Assay Kit
This Human Nrf2 assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to Nrf2 Reporter Cells, 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.
INDIGO’s cell-based reporter assays allow scientists to detect any biological activity that their test samples may exert against a specific receptor present in the cell. They utilize firefly luciferase reporter gene technology which provides superior precision and sensitivity. Since the receptor binding controls the expression of the luciferase reporter gene, luciferase activity in the cells can be correlated directly with the activity of the receptor. The strength of an interaction of a chemical with the target receptor is quantified using a luminometer to measure the level of light emitted.
Fast, reproducible, easy-to-analyze results are only four steps away
Many luciferase reporter assays require the user to grow their own cells and take time to optimize the results. INDIGO’s reporter cells contain the receptor of interest and the luciferase reporter gene. Reporter cells have been optimized to provide extreme sensitivity to quantify even small changes in receptor activity. With INDIGO’s cell-based reporter assays, the process is as easy as Thaw, Feed, Dose, and Read.
Nrf2 Reporter Cells
Nrf2 Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our cryopreserved reporter cells and assay reagents to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells are ready for immediate use so there is no need to take time on intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
Nrf2 Luciferase Reporter
INDIGO’s Nrf2 assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
Nuclear Factor (erythroid-derived 2)-like 2 Assay Services
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.
Nuclear Factor (erythroid-derived 2)-like 2 Background
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 utilizes proprietary mammalian cells that include the luciferase reporter gene functionally linked to a promoter containing tandem antioxidant response elements (AREs) and engineered to provide constitutive, high-level expression of Human Nrf2.
The principal application of this reporter assay system is in the screening of test samples to quantify functional activity, either agonist or antagonist, that may exert against Human Nrf2.
Synonyms: Nuclear Factor (erythroid-derived 2)-like 2, Nrf2, Human Nrf2, Human Nuclear Factor (erythroid-derived 2)-like 2, NFE2L2, nuclear factor erythroid-derived 2-like 2
Nuclear Factor (erythroid-derived 2)-like 2 Assay Data
Agonist dose-response analyses of Human Nrf2. Agonist analyses of Human Nrf2 Reporter Cells were performed according to the protocol described in this Technical manual, using the reference agonists CDDO Im (provided), L-Sulforaphane (Tocris), and t-BHQ (t-butyl hydroquinone; Enzo). Average values of fold-activation of Nrf2 and Z’ were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. The reference agonist t-BHQ yielded a Z' value of 0.79, confirming the robust performance of this assay and its suitability for HTS.
Nuclear Factor (erythroid-derived 2)-like 2 Assay Research Areas
Xenobiotic Metabolism; Cancer; Inflammation; Toxicology; Drug-Nutrient Interaction; Environmental Toxicology