Mouse PPARα Reporter Assay Kit

SIZE	SKU	PRICE
1 x-96 well format assays	M00111	—
3 x-32 assays in-96 well format	M00111-32	—

SIZE	SKU
1 x-96 well format assays	M00111
3 x-32 assays in-96 well format	M00111-32

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Specifications
Data
Target Background
Documentation
Overview
Citation
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Mouse Peroxisome Proliferator-Activated Receptor Alpha. INDIGO’s mPPAR alpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the mPPAR alpha. In addition to mPPAR alpha 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 mPPAR alpha.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 Type		Nuclear Hormone Receptor, Nuclear Receptor Orthologs
Species		Mouse
Receptor Form		Hybrid
Assay Mode		Agonist, Antagonist
Kit Components		Mouse PPARa Reporter Cells Cell Recovery Medium (CRM) Compound Screening Medium (CSM) GW7647, (ref. agonist; in DMSO) Detection Substrate Detection Buffer White, sterile, cell-culture ready assay plate
Shelf Life		6 months
Shipping Requirements		Dry Ice
Storage temperature		-80C

Data

Dose-response of the mouse PPARa Assay using reference agonist GW590735. Dose-response analyses of mPPARa Reporter Cells were performed according to the protocol provided in this Technical Manual. In addition, to assess the level of background signal contributed by non-specific factors that may cause activation of the luciferase reporter gene, “mock” reporter cells were prepared to contain only the luciferase vector (mock reporter cells are not provided with assay kits). mPPARa Reporter Cells and mock reporter cells were identically treated with GW590735 (Cayman Chemical) using an assay concentration range generated in 3-fold decrements: 3000, 1000, 333, 111, 37.0, 12.3, 4.12 and 0 nM. Luminescence was quantified using a GloMax-Multi+ luminometer (Promega). Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n ≥ 6). Signal-to-background (S/B) and Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. RESULTS: mPPARa reporter cells treated with 1000 nM GW590735 yielded an S/B = 118 and a corresponding Z’= 0.84. Similarly treated mock reporter cells demonstrate no significant background luminescence. Thus, luminescence results strictly through ligand-activation of the mouse PPARa expressed in these reporter cells. These data confirm the robust performance of this mPPARa Assay.

Target Background

INDIGO’s Mouse Peroxisome Proliferator-Activated Receptor Alpha (mPPARα) Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of Mouse PPARα (nr1c1), a ligand-dependent transcription factor. Because these cells incorporate a responsive luciferase reporter gene, quantifying expressed luciferase activity provides a sensitive surrogate measure of mPPARα activity in treated cells.

The primary application of this reporter assay system is in the screening of test samples to quantify functional activity, either agonist or antagonist, that they may exert against mouse PPARα.