Human TRα Reporter Assay Kit

SIZE	SKU	PRICE
1 x-96 well format assays	IB01001	—
3 x-32 assays in-96 well format	IB01001-32	—
1 x-384 well format assays	IB01002	—

SIZE	SKU
1 x-96 well format assays	IB01001
3 x-32 assays in-96 well format	IB01001-32
1 x-384 well format assays	IB01002

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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 Thyroid Hormone Receptor Alpha (TRα). INDIGO’s TR Alpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the TRα. In addition to TRα 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 TRα. 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		TRα Reporter Cells Cell Recovery Medium (CRM) Compound Screening Medium (CSM) 3,3',5-L-Triiodothyronine, (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

Activity dose-response of TRα using various reference agonists. Dose-response analyses of TRα were performed according to the protocol provided in this Technical Manual. Reporter Cells were treated with T3 (3,3’,5-L-Triiodothyronine), T2 (3,3’-L-Triiodothyronine), rT3 (3,3’,5’-L-Triiodothyronine), T4 (L-Thyroxine), KB2115, and Sobetirome. Appendix 1 in the assay technical manual provides a recommended range of treatment concentrations for T3, the reference agonist provided with this kit. Luminescence per assay well was quantified and values of average relative light units (RLU) and corresponding standard deviation (SD) were determined for each treatment concentration (n ≥ 4). Fold-activation (signal-to-background) and Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression analyses and EC50 calculations were performed using GraphPad Prism software.

Target Background

Thyroid hormone receptor alpha (TR-alpha) (erythroblastic leukemia viral (v-erb-a) oncogene homolog, avian), also known as NR1A1, is nuclear receptor protein encoded by the THRA gene. The protein encoded by this gene is a nuclear hormone receptor for triiodothyronine. It is one of the several receptors for thyroid hormone, and has been shown to mediate the biological activities of thyroid hormone. Knockout studies in mice suggest that the different receptors, while having certain extent of redundancy, may mediate different functions of thyroid hormone. Alternatively spliced transcript variants encoding distinct isoforms have been reported.

The Human TRα Reporter Assay Systems utilize proprietary human cells engineered to provide constitutive, high-level expression of Human Thyroid Hormone Receptor Alpha (NR1A1), a ligand-dependent transcription factor commonly referred to as TRα. Additionally, these cells contain a TRα-responsive luciferase reporter gene. Thus, quantifying luciferase activity provides a surrogate measure of TRα activity in the treated reporter cells.