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Mouse RORg Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays$910 USD
3 x-32 assays in-96 well format$980 USD
SIZE SKU
1 x-96 well format assays
3 x-32 assays in-96 well format

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Mouse RAR-related Orphan Receptor gamma (mRORg). INDIGO’s mRORg reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the mRORg. In addition to mRORg Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against mRORg. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeNuclear Hormone Receptor, Nuclear Receptor Orthologs
SpeciesMouse
Receptor FormHybrid
Assay ModeInverse Agonist
Kit Components
  • Mouse RORg Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Ursolic acid, (ref. inverse-agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Dose-response analyses of the mouse RORγ Assay were performed using the inverseagonists Ursolic Acid (provided),T09013173 (Tocris), and SR1001 (Cayman Chemical). Assay setup and quantification were performed as described in this Technical Manual. Relative Light Units (RLU) were quantified using a GloMax-Multi+ plate-reading luminometer (Promega Corp.). Average values of percent activity relative to the DMSO treated cells and respective values of percent coefficient of variation (%CV) were determined for each treatment concentration. Non-linear regression analyses were performed and IC50 values determined using GraphPad Prism software. Results: Ursolic acid demonstrates dose-dependent reduction of mRORγ with an IC50 ≤ 110 nM. The large assay window and corresponding high Z' value confirm the robust performance of this assay.

Target Background

RAR-related orphan receptor gamma (ROR-gamma), also known as NR1F3 (nuclear receptor subfamily 1, group F, member 3) 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 Mouse RORγ (RAR-related Orphan Receptor Gamma; nr1f3) referred to as mouse rorC, or here as mRORγ. The N-terminal DNA binding domain (DBD) of the native mRORγ receptor has been substituted with that of the yeast GAL4-DBD. The principle application of this reporter assay is in the screening of test samples to quantify inverse-agonist or agonist activities that they may exert against mouse RORγ.

Also available as a service

RAR-related Orphan Receptor Gamma (RORg, NR1F3)

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