RAR-related Orphan Receptor Gamma (RORγ; NR1F3)
|Product Family||Product Number||Product Description||Technical Manual|
|IB04001-32||Human RORγ Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB04001||Human RORγ Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB04002||Human RORγ Reporter Assay System, 1 x 384-well format assays|
RAR-related Orphan Receptor Gamma Assay Kit
This RAR-related Orphan Receptor Gamma (RORγ) assay kit is an all-inclusive firefly luciferase reporter assay system that includes, in addition to RORγ Reporter Cells, two optimized media for use during cell culture and (optionally) in diluting the test samples, a reference inverse-agonist, Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol.
INDIGO’s cell-based reporter assays allow scientists to detect any biological activity that their test samples may exert against a specific receptor present in the cell. They utilize firefly luciferase reporter gene technology which provides superior precision and sensitivity. Since the receptor binding controls the expression of the luciferase reporter gene, luciferase activity in the cells can be correlated directly with the activity of the receptor. The strength of an interaction of a chemical with the target receptor is quantified using a luminometer to measure the level of light emitted.
Fast, reproducible, easy-to-analyze results are only four steps away
Many luciferase reporter assays require the user to grow their own cells and take time to optimize the results. INDIGO’s reporter cells contain the receptor of interest and the luciferase reporter gene. Reporter cells have been optimized to provide extreme sensitivity to quantify even small changes in receptor activity. With INDIGO’s cell-based reporter assays, the process is as easy as Thaw, Feed, Dose, and Read.
RORγ Reporter Cells
RORγ Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our cryopreserved reporter cells and assay reagents to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells are ready for immediate use so there is no need to take time on intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
RORγ Luciferase Reporter
INDIGO’s RORγ 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.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
RORg assay kits also available in: Mouse
RAR-related Orphan 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.
RORg Service Assays: Human, Mouse
RAR-related Orphan Receptor Gamma Background
RAR-related orphan receptor gamma (ROR-gamma), also known as NR1F3 is a nuclear receptor encoded by the RORG gene.The RAR-related orphan receptors (RORs) are members of the nuclear receptor family of intracellular transcription factors. There are three forms of ROR, ROR-α, -β, and -γ and each is encoded by a separate gene (RORA, RORB, and RORC respectively). The RORs are somewhat unusual in that they appear to bind as monomers to hormone response elements as opposed to the majority of other nuclear receptors which bind as dimers. Melatonin has been reported to be the endogenous ligand for ROR-α while CGP 52608 has been identified as a ROR-α selective synthetic ligand. However X-ray crystallographic (PDB 1n83 and 1s0x) and functional data both suggest that cholesterol or a cholesterol derivative may be the endogenous ligand. In contrast, all-trans retinoic acid binds with high affinity to ROR-β and -γ but not ROR-α.
INDIGO’s Human RAR-related Orphan Receptor Gamma (NR1F3; RORγ) Assay utilizes proprietary human cells engineered to provide high-level expression of a hybrid form of RORγ. The N-terminal DNA binding domains (DBD) of the native RORγ and RORγt receptors (isoforms 1 and 2, respectively) have been substituted with that of the yeast GAL4-DBD. Hence, the GAL4-RORγ hybrid expressed in these reporter cells will not discern functional differences that may exist between the native isoform 1 and isoform 2 receptors. Endogenous molecular activators maintain RORγ in a state of constitutive high-level activity. Therefore, the principle application of this reporter assay system is in the screening of test samples to quantify inverse-agonist or agonist activities that they may exert against human RORγ.
For more information on RORγ, visit the Nuclear Receptor Resource.
RAR-related Orphan Receptor Gamma Assay Data
The Human RORγ Assay. Dose-response analyses of the RORγ Assay were performed using the inverse-agonists ML-209 (provided as reference), Ursolic Acid, AZ 5104, TO901317, and SR 1001 (all from Cayman Chemical, USA). RORγ Assay setup was performed as described in the assay Technical Manual. Relative Light Units were quantified and average values of RLU and their respective values of relative standard deviation were determined for each treatment concentration. Non-linear regression analyses were performed and IC50 values determined using GraphPad Prism software. The high Z' value confirms the robust performance of this assay, and its suitability for HTS applications.
RAR-related Orphan Receptor Gamma Assay Research Areas
Inflammation; CNS, Circadian and Basal Metabolism; Bile Acid and Xenobiotic; Retinoid Panel; NASH/NAFLD