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    • Assay Kit Platform & Formats
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    • Animal Model Assays
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    • Live Cell Multiplex
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Zebrafish Peroxisome Proliferator-Activated Receptor Gamma (zPPARγ; nr1c3)

Product Family Product Number Product Description Technical Manual
Z0010
zPPARγ
(nr1c3)
Z00101-32 Zebrafish PPARγ Reporter Assay System, 3 x 32 assays in 96-well format Technical Manual
Z00101 Zebrafish PPARγ Reporter Assay System, 1 x 96-well format assays Technical Manual

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  • Kits
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  • Background
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Zebrafish Peroxisome Proliferator-Activated Receptor Gamma Assay Kit INDIGO Assay Kit

This Zebrafish Peroxisome Proliferator Activated Receptor Gamma (zPPARγ) assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to zPPARγ Reporter Cells, two optimized media for use during cell culture and (optionally) in diluting the test samples, a reference 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.

Thaw, Feed, Dose, Read

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.

Zebrafish PPARγ Reporter Cells

Zebrafish PPARγ 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.

Zebrafish PPARγ Luciferase Reporter

INDIGO’s Zebrafish PPARγ 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.

Assay Kit & Platforms

Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.

PPARg assay kits also available in: Human, Rat/Mouse Shared Ortholog, Cyn Monkey

Zebrafish Peroxisome Proliferator-Activated Receptor Gamma Assay Services

The primary application of INDIGO’s cell-based Zebrafish PPARγ assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of Zebrafish PPARγ. 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.

Service Assays: Human, Rat/Mouse Shared Ortholog, Cyn Monkey, Zebrafish

Zebrafish Peroxisome Proliferator-Activated Receptor Gamma Assay 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.

For more information on PPARγ, visit the Nuclear Receptor Resource.

Zebrafish Peroxisome Proliferator-Activated Receptor Gamma Assay Research Areas

Diabetes; Obesity; Cancer; Inflammation; Lipid Metabolism and Energy; Metabolic Disease; NASH/NAFLD; Environmental Toxicology

Defining the Chemical Additives Driving In Vitro Toxicities of Plastics

ABSTRACT Increases in the global use of plastics have caused concerns regarding potential adverse effects on human health. Plastic products contain hundreds of potentially toxic chemical additives, yet the exact chemicals which drive toxicity currently remain unknown. In this study, we employed nontargeted analysis and in vitro bioassays to identify the toxicity drivers in plastics.Read More

Posted in New Publications | Tagged PPARγ | Comments Off on Defining the Chemical Additives Driving In Vitro Toxicities of Plastics

Lipoxygenase catalyzed metabolites derived from docosahexaenoic acid are promising antitumor agents against breast cancer

ABSTRACT Docosahexaenoic acid (DHA) is known to inhibit breast cancer in the rat. Here we investigated whether DHA itself or select metabolites can account for its antitumor action. We focused on metabolites derived from the lipoxygenase (LOX) pathway since we previously showed that they were superior anti-proliferating agents compared to DHA; 4-OXO-DHA was the mostRead More

Posted in New Publications | Tagged PPARg, PPARγ | Comments Off on Lipoxygenase catalyzed metabolites derived from docosahexaenoic acid are promising antitumor agents against breast cancer

Nuclear Receptors Regulate Intestinal Inflammation in the Context of IBD

ABSTRACT Gastrointestinal (GI) homeostasis is strongly dependent on nuclear receptor (NR) functions. They play a variety of roles ranging from nutrient uptake, sensing of microbial metabolites, regulation of epithelial intestinal cell integrity to shaping of the intestinal immune cell repertoire. Several NRs are associated with GI pathologies; therefore, systematic analysis of NR biology, the underlyingRead More

Posted in New Publications | Tagged glucocorticoids, GR, ibd, Inflammation, intestinal inflammation, nuclear receptors, PPARγ, PXR, VDR | Comments Off on Nuclear Receptors Regulate Intestinal Inflammation in the Context of IBD

Structures and biological activities of new carnosic acid- and carnosol-related compounds generated by heat treatment of rosemary

ABSTRACTCarnosic acid (CA) and carnosol (CS) are components of rosemary reported to possess antioxidative and peroxisome proliferator-activated receptor gamma (PPARγ) transcriptional activities that are useful for preventing metabolic disorders. The aims of this study were to identify new bioactive compounds in heated rosemary and examine their effects on antioxidative stress and PPARγ activation. In heatedRead More

Posted in New Publications | Tagged Carnosic acid, carnosol, PPAR gamma, PPARγ | Comments Off on Structures and biological activities of new carnosic acid- and carnosol-related compounds generated by heat treatment of rosemary

Tetrahydro-Isohumulone Derivatives, Methods of Making and Using: Patent Application

ABSTRACT The present application provides novel tetrahydro – isohumulone ( THIAA ) derivatives and substantially enantiomerically pure compositions and pharmaceutical formulations thereof . The application further provides methods of using the disclosed compounds and compositions to activate PPARy , inhibit inflammation , and treat conditions associated with inflammation and conditions responsive to PPARY modulation suchRead More

Posted in New Publications | Tagged patent, PPAR, PPARγ | Comments Off on Tetrahydro-Isohumulone Derivatives, Methods of Making and Using: Patent Application

Comparative Evaluation of Gemcabene and Peroxisome Proliferator–Activated Receptor Ligands in Transcriptional Assays of Peroxisome Proliferator–Activated Receptors: Implication for the Treatment of Hyperlipidemia and Cardiovascular Disease

ABSTRACT Gemcabene, a late-stage clinical candidate, has shown efficacy for LDL-C, non-HDL cholesterol, apoB, triglycerides, and hsCRP reduction, all risk factors for cardiovascular disease. In rodents, gemcabene showed changes in targets, including apoC-III, apoA-I, peroxisomal enzymes, considered regulated through peroxisome proliferator–activated receptor (PPAR) gene activation, suggesting a PPAR-mediated mechanism of action for the observed hypolipidemic effects observedRead More

Posted in New Publications | Tagged gemcabene, nuclear hormone receptors, PPARα, PPARγ, PPARδ, transactivation | Comments Off on Comparative Evaluation of Gemcabene and Peroxisome Proliferator–Activated Receptor Ligands in Transcriptional Assays of Peroxisome Proliferator–Activated Receptors: Implication for the Treatment of Hyperlipidemia and Cardiovascular Disease

GPR40 partial agonist MK-2305 lower fasting glucose in the Goto Kakizaki rat via suppression of endogenous glucose production

ABSTRACT GPR40 (FFA1) is a fatty acid receptor whose activation results in potent glucose lowering and insulinotropic effects in vivo. Several reports illustrate that GPR40 agonists exert glucose lowering in diabetic humans. To assess the mechanisms by which GPR40 partial agonists improve glucose homeostasis, we evaluated the effects of MK-2305, a potent and selective partialRead More

Posted in New Publications | Tagged glucose metabolism, glucose stimulated insulin secretion, ortholog, PPAR, PPARα, PPARβ, PPARγ, rat PPAR | Comments Off on GPR40 partial agonist MK-2305 lower fasting glucose in the Goto Kakizaki rat via suppression of endogenous glucose production

Structural basis for differential activities of enantiomeric PPARγ agonists: Binding of S35 to the alternative site

ABSTRACT Peroxisome proliferator-activated receptor γ (PPARγ) is a member of the nuclear receptor superfamily. It functions as a ligand-activated transcription factor and plays important roles in the regulation of adipocyte differentiation, type 2 diabetes mellitus, and inflammation. Many PPARγ agonists bind to the canonical ligand-binding pocket near the activation function-2 (AF-2) helix (i.e., helix H12)Read More

Posted in New Publications | Tagged enantiomeric PPARγ, PPAR, PPARγ, PPARγ agonist | Comments Off on Structural basis for differential activities of enantiomeric PPARγ agonists: Binding of S35 to the alternative site

Pyrintegrin Induces Soft Tissue Formation by Transplanted or Endogenous Cells

ABSTRACT Focal adipose deficiency, such as lipoatrophy, lumpectomy or facial trauma, is a formidable challenge in reconstructive medicine, and yet scarcely investigated in experimental studies. Here, we report that Pyrintegrin (Ptn), a 2,4-disubstituted pyrimidine known to promote embryonic stem cells survival, is robustly adipogenic and induces postnatal adipose tissue formation in vivo of transplanted adiposeRead More

Posted in New Publications | Tagged PPAR, PPARγ | Comments Off on Pyrintegrin Induces Soft Tissue Formation by Transplanted or Endogenous Cells

Lesinurad, a novel, oral compound for gout, acts to decrease serum uric acid through inhibition of urate transporters in the kidney

ABSTRACT Background Excess body burden of uric acid promotes gout. Diminished renal clearance of uric acid causes hyperuricemia in most patients with gout, and the renal urate transporter (URAT)1 is important for regulation of serum uric acid (sUA) levels. The URAT1 inhibitors probenecid and benzbromarone are used as gout therapies; however, their use is limitedRead More

Posted in New Publications | Tagged PPAR, PPARγ | Comments Off on Lesinurad, a novel, oral compound for gout, acts to decrease serum uric acid through inhibition of urate transporters in the kidney

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