Ortholog Assays: Maximize the Potential of Animal Studies

INDIGO Biosciences offers 40+ ortholog assays to streamline animal studies and aid drug development.

Facilitate cross-species comparisons in animal studies

Select the most suitable animal model as a surrogate for humans

Enable quicker assessment with confident data interpretation


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Cross Species Comparisons with Ortholog Assays

While animal studies are widely used for evaluating chemical interactions with our biological systems, they present numerous challenges that significantly impede the development of drug candidates and can cause delays in moving to clinical trials. Recognizing these issues and the lack of solutions, INDIGO Biosciences has developed and validated over 40 ortholog nuclear receptor assays for a variety of commonly used laboratory animals such as rats, mice, dogs, monkeys, rabbits, and zebrafish.

By offering this unique portfolio of ortholog assays, INDIGO Biosciences facilitates cross-species comparisons, enabling researchers to effectively interpret the results obtained from previous animal trials or to select the most appropriate animal model as a surrogate for humans. Now, it is possible to assess differences in the activities of ortholog receptors compared to their human counterparts within a matter of days, allowing for quicker evaluation of drug-specific effects.

With our comprehensive selection of ortholog assays, you can confidently screen the appropriate animal model prior to conducting trials and interpret your data with certainty. Don't leave anything to chance.

Frequently Asked Questions

INDIGO’s assays are not binding assays. They are cell-based trans-activation assays, and the principal application is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that the compounds may exert against the nuclear receptors. INDIGO reporter systems utilize firefly luciferase reporter gene technology, and while there is a binding taking place, our assays do not measure it. Instead, the luciferase light response is measured which correlates to the activation status of the receptor (either activation or inhibition). Quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in receptor activity. It will tell the scientist the significance (strength) of the interaction by the level of light emitted. In addition, cell-based assays are more sensitive and able to detect smaller levels of activation.

INDIGO’s nuclear receptor assays utilize proprietary human and non-human mammalian cells engineered to provide constitutive, high-level expression of the designated receptor. Specific cell type information for each assay is proprietary and available only through consultation with INDIGO’s technical team following a screening service or assay kit purchase.

Reporter cells included in INIDIGO’s steroid hormone nuclear assay kits (ERα, ERβ, AR, PGR, MR, GR) express the native, full-length receptors. INDIGO’s other nuclear receptor assays, however, include reporter cells that express hybrid nuclear receptors. In these cases, the respective receptor’s native N-terminal sequence comprising the DNA Binding Domain (DBD) has been replaced with sequence encoding the yeast Gal4-DBD. All other native NR functional/structural domains (ligand binding domain, hinge region, and various activation domains) are present in these hybrid receptors. These reporter cells also contain the firefly luciferase reporter gene functionally linked to the upstream genetic response element for Gal4. Consequently, once a bioactive compound associates with the ligand binding domain of the hybrid receptor, only the luciferase reporter gene is induced. Ligand-activation of the hybrid receptor will not induce collateral expression of target genes that are otherwise regulated by the native nuclear receptor.

View our list of reference compounds. We do utilize commercially available reference agonists for our assays, and this reference agonist is included in each assay kit. We do use reference antagonists where a reference is known and commercially available. Some of our antagonist assays do not have reference antagonists available.

Each kit includes luciferase reporter cells, an assay plate, all required reagents (optimized culture media, positive-control agonist, and detection reagent), and a detailed Technical Manual. A luminometer is required for Relative Light Unit (RLU) measurements.

INDIGO’s kits are intended for research purposes only, and not for diagnostic or therapeutic use in humans.

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