Zebrafish Retinoic Acid Receptor Alpha isoform A (zRARαa; nr1b1 isoform A)
|Product Family||Product Number||Product Description||Technical Manual|
(nr1b1 iso A)
|Z02201-32||Zebrafish RARαA Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|Z02201||Zebrafish RARαA Reporter Assay System, 1 x 96-well format assays||Technical Manual|
Zebrafish Retinoic Acid Receptor Alpha A Assay Kit
This Zebrafish Retinoic Acid Receptor Alpha isoform A (zRARαa)assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to zRARαa 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.
zRARαa 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 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./p>
Kits are offered in different assay formats to accommodate researchers’ needs: 3x 32 and 1x 96, 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.
Zebrafish Retinoic Acid Receptor Alpha A 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.
Zebrafish Retinoic Acid Receptor Alpha A Assay 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.
INDIGO’s Zebrafish Retinoic Acid Receptor, Alpha A (zRARαa) Reporter Assay System utilizes proprietary non-human cells engineered to provide constitutive, high-level expression of Zebrafish (Danio rerio) RARαa (NR1B1 isoform A), a ligand-dependent transcription factor.
INDIGO's Reporter Cells express a hybrid zebrafish RARαa receptor in which the native N-terminal ligand binding domain (LBD) has been substituted with that of the yeast GAL4 LBD sequence. Accordingly the resident luciferase reporter gene is functionally linked to tandem copies of the Gal4 upstream activation sequence. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of changes in zRARαa. The principal application of this assay is in the screening of test samples to quantify any functional bioactivity that they may exert against zebrafish RARαa. In particular, zebrafish reporter assays are frequently used in the monitoring of environmental samples for the presence of biohazardous chemical pollutants, such as endocrine disruptors.
For more information on RARα, visit the Nuclear Receptor Resource.