Human TRβ Reporter Assay Kit

SIZE	SKU	PRICE
1 x-96 well format assays	IB01101	—
3 x-32 assays in-96 well format	IB01101-32	—
1 x-384 well format assays	IB01102	—

SIZE	SKU
1 x-96 well format assays	IB01101
3 x-32 assays in-96 well format	IB01101-32
1 x-384 well format assays	IB01102

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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 Beta (TRβ). INDIGO’s TR Beta reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the TRβ. In addition to TRβ 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 TRβ. 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		TRβ 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		Yes
Shipping Requirements		Dry Ice
Storage temperature		-80C

Data

Activity dose-response of TRβ using various reference agonists. Dose-response analyses of TRβ 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

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. Mutations in this gene are known to be a cause of generalized thyroid hormone resistance (GTHR), a syndrome characterized by goiter and high levels of circulating thyroid hormone (T3-T4), with normal or slightly elevated thyroid stimulating hormone (TSH). Several alternatively spliced transcript variants encoding the same protein have been observed for this gene.

INDIGO’s TRβ Reporter Assay System utilizes proprietary human cells engineered to provide constitutive, high-level expression of Human Thyroid Hormone Receptor Beta (NR1A2), a ligand-dependent transcription factor. Because these cells incorporate a responsive luciferase reporter gene, quantifying expressed luciferase activity provides a sensitive surrogate measure of TRβ activity in the treated cells.

The primary 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 TRβ.