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Zebrafish PPARg 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 Zebrafish Peroxisome Proliferator-Activated Receptor gamma. INDIGO’s zPPAR gamma reporter assay utilizes proprietary non-human cells that have been engineered to provide constitutive expression of the zPPAR gamma. In addition to zPPAR gamma 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 zPPAR gamma. 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
SpeciesZebrafish
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Zebrafish PPARg Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Rosiglitazone, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Agonist dose-response analyses of the Zebrafish PPARg Assay. Validation of the zfPPARg Assay was performed following the protocol described in the Technical Manual, using the reference agonists Triphenyl Phosphate (TPP; provided), Troglitazone, and Mono-2-ethylhexyl phthalate (MEHP). zfPPAR Reporter Cells were treated with concentrations of TPP as described in Appendix 1. Luminescence was quantified using a plate-reading luminometer. Values of average Relative Light Units (RLU; average of n = 4), standard deviation (SD), Fold-Activation and Coefficient of Variation (%CV) were determined. Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression analyses were performed and EC50 values determined using GraphPad Prism software.

Target Background

Peroxisome proliferator-activated receptor gamma (PPAR-gamma or PPARγ), also known as the glitazone receptor, or NR1C3 (nuclear receptor subfamily 1, group C, member 3) is a type II nuclear receptor that in humans is encoded by the PPARG gene. PPARs form heterodimers with retinoid X receptors (RXRs) and these heterodimers regulate transcription of various genes. PPARγ regulates adipocyte differentiation, fatty acid storage and glucose metabolism. The PPARγ knockout mice fail to generate adipose tissue when fed a high fat diet. Many insulin sensitizing drugs used in the treatment of diabetes target PPARγ as a means to lower serum glucose without increasing pancreatic insulin secretion. Additionally, PPARγ has been implicated in the pathology of numerous diseases including obesity, diabetes, atherosclerosis and cancer. Alternatively spliced transcript variants that encode different isoforms have been described.

INDIGO’s Zebrafish Peroxisome Proliferator Activated Receptor Gamma Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of Zebrafish (Danio rerio) PPARγ (nr1c3), a ligand-dependent transcription factor.

INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to an zPPARγ-responsive promoter. Thus, quantifying changes in luciferase expression in the test sample-treated reporter cells provides a sensitive surrogate measure of changes in zPPARγ activity. The principal application of this assay is in the screening of test samples to quantify any functional bioactivity that they may exert against zebrafish PPARγ. 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.

Also available as a service

Peroxisome Proliferator-Activated Receptor Gamma (PPARg, NR1C3)

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