Human RORα Reporter Assay Kit

SIZE	SKU	PRICE
1 x-96 well format assays	IB04011	—
3 x-32 assays in-96 well format	IB04011-32	—
1 x-384 well format assays	IB04012	—

SIZE	SKU
1 x-96 well format assays	IB04011
3 x-32 assays in-96 well format	IB04011-32
1 x-384 well format assays	IB04012

Add to quote

Specifications
Data
Target Background
Documentation
Overview
Citation
Related Products

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human RAR-related Orphan Receptor Alpha (RORa). INDIGO’s ROR Alpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the ROR Alpha. In addition to ROR Alpha Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human ROR Alpha. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

Ready to Use Upon Receipt
Includes All Needed Components
Contains Transfected Reporter Cells
Eliminates Cell Licensing Fees
Clear, Reproducible Results
Consistent Results Lot to Lot

Product Specifications

Target Type		Nuclear Hormone Receptor
Species		Human
Receptor Form		Hybrid
Assay Mode		Inverse Agonist
Kit Components		RORa Reporter Cells Cell Recovery Medium (CRM) Compound Screening Medium (CSM) All trans Retinoic Acid, (ref. inverse-agonist; in DMSO) Detection Substrate Detection Buffer White, sterile, cell-culture ready assay plate
Shelf Life		6 months
Shipping Requirements		Dry Ice
Storage temperature		-80C

Data

Agonist dose-response performance of the Human RORα assay. Agonist response of the Human RORα Reporter Cells is demonstrated using 27-Hydroxy Cholesterol (27OHC; Cayman Chemical), 7-dehydro Cholesterol (Sigma), and CGP 52608 (Sigma). Values of Fold-Activation are plotted against concentration. 27OHC provides greater than a 2-fold increase in RORα activity above the already high endogenous activity level. Z' values confirm the robust performance of the agonist-mode RORα assay. When contemplating concentration ranges for screening test compounds of unknown bioactivity, it is important to note the great disparity in potencies between inverse-agonists and theses agonist reference compounds: ATRA IC50 1 µM.

Inverse-agonist dose-response performance of the Human RORα assay. Inverse-agonist analyses of Human RORα Reporter Cells using All trans-Retinoic Acid (ATRA; provided). Luminescence was quantified using a GloMax-Multi+ luminometer (Promega). Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n ≥ 6). Z’ value was calculated as described by Zhang, et al. (1999). Non-linear regression was performed using GraphPad Prism software.

Target Background

RAR-related orphan receptor alpha (ROR-alpha), also known as NR1F1 is a nuclear receptor encoded by the RORA gene. The protein encoded by this gene is a member of the NR1 subfamily of nuclear hormone receptors. It can bind as a monomer or as a homodimer to hormone response elements upstream of several genes to enhance the expression of those genes. The specific functions of this protein are not known, but it has been shown to interact with NM23-2, a nucleoside diphosphate kinase involved in organogenesis and differentiation, as well as with NM23-1, the product of a tumor metastasis suppressor candidate gene. Four transcript variants encoding different isoforms have been described for this gene.

INDIGO’s RAR-related Orphan Receptor Alpha (RORα) assay includes proprietary mammalian cells engineered to provide high-level expression of Human RORα. Human RAR Orphan Receptor Alpha Inverse Agonist Assay Reporter Cells also incorporate a responsive luciferase reporter gene. Quantifying expressed luciferase activity at the assay endpoint provides a sensitive surrogate measure of changes in RORα activity in treated cells. Endogenous molecular activators maintain RORα in a state of constitutive high-level activity. Therefore, the principle application of this reporter assay system is in the screening of test samples to quantify inverse-agonist or agonist activities that they may exert against human RORα.