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Biology Chemistry DMPK Pharmacology Toxicology

TRalpha Reporter Assay

Service Description

Provider: INDIGO Biosciences, Inc. provides Nuclear Receptor Reporter Assay based products and services for drug discovery and toxicology. INDIGO specializes in the development of frozen, whole-cell Nuclear Receptor Assay Systems for use in small compound HTS, off-target drug interactions, and toxicology screens.

Background: Thyroid hormone receptor alpha (TR-alpha or TRα), also known as NR1A1 (nuclear receptor subfamily 1, group A, member 1), is a nuclear receptor encoded by the THRA gene (erythroblastic leukemia viral (v-erb-a) oncogene homolog, avian). TRα is one of the several receptors for thyroid hormone, triiodothyronine, 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.

Service Details: This assay can be utilized to examine the agonism (activation) or antagonism (inhibition of an agonist) on the nuclear receptor. Mammalian cells are transfected with the ligand binding domain of the NR fused to a heterologous DNA binding domain as well as the response element for this DNA binding motif linked to a reporter gene. A transfection efficiency control plasmid is also included. These transfected cells are seeded in 96-well plates and then treated overnight with test compound at 37°C. Cells are then lysed and NR activation reporter activity as well as transfection efficiency reporter activity is assessed. A positive control compound (6 concentrations in triplicate) as well as vehicle control (triplicate) is included in every experiment.

For dose response curves, it is suggested that at least six concentrations of the test compounds be examined, covering a wide range of dilutions (3 log-units). In addition, we recommend that each compound/concentration be assayed in triplicate.

Deliverable: Downloadable data report containing raw and analyzed data. EC~50~ and peak activity will be determined using nonlinear regression. Dose-response data will be plotted for the test compounds as well as the positive control. A sample data report is available on request.

Price: The listed price ($1000) is the cost of assay set-up fee. In addition, there is a $15 per well charge (one compound assayed at one concentration). The per well price is $10 if the sample size is greater than 960 wells. The final price is calculated based on the number of wells to be tested. For example, to compare the activity of two compounds to that of a well-known agonist of this receptor, six concentrations of each compound as well as a vehicle reference would be examined. This would be a total of 57 wells and the cost of the experiment is $1855.

Sample Submission: Enter sample submission information and select service options in the Service Order Form that you can access once you add this item to your cart. The minimum sample requirement is 25 µl of a 10 mM stock solution, with the preferred sample submission of 100 mM stock solution in DMSO.

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Click on the orange 'Add to Cart' button to access the Service Order Form, which contains technical questions and sample submission information. The form allows you to type in or paste in information or to select from a list of pre-defined choices. Complete and submit the Service Order Form to start the checkout process.

Once you submit your order, you can communicate with the provider or upload files by using Assay Depot’s microblogging system. The microblogging system allows you to stay in direct contact with the provider the entire time a research service is being carried out.


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