Human RXRα Reporter Assay Kit

SIZE	SKU	PRICE
1 x-96 well format assays	IB00801	—
3 x-32 assays in-96 well format	IB00801-32	—
1 x-384 well format assays	IB00802	—

SIZE	SKU
1 x-96 well format assays	IB00801
3 x-32 assays in-96 well format	IB00801-32
1 x-384 well format assays	IB00802

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Specifications
Data
Target Background
Documentation
Overview
Citation
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Retinoid X Receptor alpha (RXRa). INDIGO’s RXR Alpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the RXR Alpha. In addition to RXR 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 RXR 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		Agonist, Antagonist
Kit Components		RXRa Reporter Cells Cell Recovery Medium (CRM) Compound Screening Medium (CSM) 9-cis Retinoic Acid, (ref. agonist; in DMSO) Detection Substrate Detection Buffer White, sterile, cell-culture ready assay plate
Shelf Life		6 months
Orthologs Available		No
Shipping Requirements		Dry Ice
Storage temperature		-80C

Data

Agonist and Antagonist dose-response analyses of the Human RXRα. A.) Analyses of RXRα Reporter Cells using the reference agonists 9-cis-Retinoic Acid (provided). B.) Analyses of RXRα antagonist dose-responses using HX531 and UVI3003 (Tocris). Assay setups and quantification of RXRα activity were performed following the protocol provided in the assay Technical Manual. Luminescence was quantified and average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n ≥ 4). Values of Fold-activation and Z’ were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. High S/B and Z' scores confirm the robust performance of this RXRα Assay.

Target Background

Retinoid X receptor alpha (RXRα), also known as NR2B1 is a nuclear receptor which in humans is encoded by the RXRA gene. Retinoid X receptors (RXRs) and retinoic acid receptors (RARs), are nuclear receptors that mediate the biological effects of retinoids by their involvement in retinoic acid-mediated gene activation. These receptors exert their action by binding, as homodimers or heterodimers, to specific sequences in the promoters of target genes and regulating their transcription. The protein encoded by this gene is a member of the steroid and thyroid hormone receptor superfamily of transcription factors.

INDIGO’s RXR Alpha Reporter Assay System utilizes mammalian cells engineered to provide constitutive, high-level expression of Human Retinoid X Receptor Alpha (NR2B1), a ligand-dependent transcription factor commonly referred to as RXRA or RXRα. Additionally, these cells contain an RXRα-responsive luciferase reporter gene. Thus, quantifying luciferase activity provides a surrogate measure of RXRα activity in the treated reporter cells.