Retinoic Acid Receptor Gamma (RARγ; NR1B3)
| Product Family | Product Number | Product Description | Technical Manual |
| IB0200 RARγ (NR1B3) |
IB02001-32 | Human RARγ Reporter Assay System, 3 x 32 assays in 96-well format | Technical Manual |
| IB02001 | Human RARγ Reporter Assay System, 1 x 96-well format assays | Technical Manual | |
| IB02002 | Human RARγ Reporter Assay System, 1 x 384-well format assays | Technical Manual |
Retinoic Acid Receptor Gamma Assay Kit
This Retinoic Acid Receptor Gamma (RARγ assay kit is an all-inclusive RAR gamma reporter assay system that includes, in addition to RARγ Reporter Cells, two optimized media for use during cell culture and (optionally) in diluting the test samples, a reference agonist, Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol.
RARγ Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
INDIGO’s RARγ assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
RARγ reporter assay kits are offered in different assay formats to accommodate researchers’ needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
Retinoic Acid Receptor Gamma Assay Services
The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. Service assays include a positive control reference compound and ‘vehicle’ control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online “Request a Quote” form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.
Retinoic Acid Receptor Gamma Background
Retinoic acid receptors (RARs) are nuclear hormone receptors of the NRB1 class, which function as heterodimers with retinoid X receptors (RXRs). There are three distinct RAR subtypes; RARalpha, RARbeta and RARgamma. RARalpha is present in most tissue types, whereas RARbeta and RARgamma expression is more selective. RXR-RAR heterodimers act as ligand-dependent transcriptional regulators by binding to the specific retinoic acid response element (RARE) found in the promoter regions of target genes. In the absence of an RAR agonist, RXR-RAR recruits co-repressor proteins such as NCoR and associated factors such as histone deacetylase to maintain a condensed chromatin structure. RAR agonist binding stimulates co-repressor release and co-activator complexes, such as histone acetyltransferase, are recruited to activate transcription. RARs transduce retinoid signals in vivo, which mediates proper embryogenesis, differentiation and growth arrest. Specifically, RXRalpha-RARgamma heterodimers are necessary for growth arrest and viseral and primitive endodermal differentiation, whereas RXRalpha-RARalpha is required for cAMP-dependent parietal endodermal differentiation. In vitro it has been difficult to elucidate the roles of individual subtypes as functional RAR knockouts generate artificial redundancies that are thought not to exist under normal conditions.
The Human RARγ Reporter Assay Systems utilize proprietary mammalian cells engineered to provide constitutive, high-level expression of Human Retinoic Acid Receptor Gamma (NR1B3), a ligand-dependent transcription factor commonly referred to as RARγ. Additionally, these cells contain a RARγ-responsive luciferase reporter gene. Thus, quantifying luciferase activity provides a surrogate measure of RARg activity in the treated reporter cells.
For more information on RARγ, visit the Nuclear Receptor Resource.
Synonyms: Retinoic Acid Receptor Gamma, RAR gamma, RARγ, RARg, Human Retinoic Acid Receptor Gamma, NR1B3
Retinoic Acid Receptor Gamma Data
Agonist dose-response of the RARγ Assay. Validation of the RARγ Assay was performed using manual dispensing and following the protocol described in this Technical Manual, using the reference agonists all-trans-Retinoic Acid (provided), Adapalene, BMS 961, and CD1530 (all from Tocris). In addition, to assess the level of background signal contributed by non-specific factor(s) that may cause activation of the luciferase reporter gene, “mock” reporter cells were specially prepared to contain only the luciferase reporter vector (mock reporter cells are not provided with assay kits). RARγ Reporter Cells and Mock reporter cells were identically treated with trans-retinoic acid. Luminescence was quantified using a GloMax-Multi+ plate-reading luminometer (Promega Corp.). Average relative light units (RLU) and respective standard deviation (SD) and Signal-to-Background (S/B) values were determined for each treatment concentration (n ≥ 6). Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression analyses were performed and EC50 values determined using GraphPad Prism software. Results: Mock reporter cells treated with trans-retinoic acid demonstrate no significant background luminescence (≤ 0.05% that of the reporter cells at ECMax). Thus, luminescence results strictly through ligand-activation of the human RARγ expressed in these reporter cells. These data confirm the robust performance of this RARγ assay and demonstrate its suitability for use in HTS applications.
Validation of RARγ Assay antagonist dose-responses. RARγ antagonist assays were performed using MM11253, CD2665, BMS453 and ER50891 (all from Tocris). Assay setup and quantification of RARγ activity were performed following the protocol described in this Technical Manual. Final assay concentrations of the respective antagonists ranged between 10 µM and 10 pM, and included a 'no antagonist' control (n ≥ 6 per treatment; highest [DMSO] ≤ 0.1% f.c.). Each treatment also contained 3.8 nM (~ EC80) of trans-Retinoic Acid. Assay plates were incubated for ~24 hrs, then processed to quantify RARγ activity for each treatment condition.
Retinoic Acid Receptor Gamma Assay Research Areas
Cancer; Autoimmune Disease; Psoriasis; Reproduction and Development; Retinoid Panel