The panel of PPAR reporter assays utilizes non-human mammalian cells engineered to express human peroxisome proliferator-activated receptors: PPARα (NR1C1), PPARδ (NR1C2), or PPARγ (NR1C3), all ligand-dependent transcription factors that are commonly referred to as PPARα, PPARδ and PPARγ. The PPAR reporter cells include the luciferase reporter gene functionally linked to a responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in PPARα, PPARδ, or PPARγ activity. The principal application of this assay panel is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against the three human PPARs. PPAR reporter cells are prepared using the proprietary CryoMite process. This cryo-preservation method yields exceptional cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for cumbersome intermediate treatment steps such as spin-and- rinse of cells, viability determinations, cell titer adjustments, or the pre-incubation of reporter cells prior to assay setup. The nuclear receptor reporter assays are all-inclusive cell-based assay systems. In addition to PPAR 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.
▪ PPARα Reporter Cells: 1 x 0.60 mL; -80°C ▪ PPARδ Reporter Cells: 1 x 0.60 mL; -80°C ▪ PPARγ Reporter Cells: 1 x 0.60 mL; -80°C▪ Cell Recovery Media (CRM): 1 x 10.5 mL; -20°C or +4°C (< 5 days)▪ Compound Screening Media (CSM): 1 x 35 mL; -20°C or +4°C (< 5 days) ▪ PPARα reference agonist: GW590735, 10 mM (in DMSO): 1 x 30 µL; -20°C ▪ PPARδ reference agonist: GW0742, 1.0 mM (in DMSO): 1 x 30 µL; -20°C ▪ PPARγ reference agonist: Rosiglitazone, 10 mM (in DMSO): 1 x 30 µL; -20°C▪ Detection Substrate: 3 x 2.0 mL; -80°C▪ Detection Buffer: 3 x 2.0 mL; -80°C ▪ 96-well format plate frame: 1; ambient ▪ snap-in, 8-well strips (white, sterile, cell culture treated): 12; ambient