Zebrafish Assay Solutions
Zebrafish are becoming an increasingly popular model for environmental toxicology, as well as developmental and biological research as their easy maintenance, high reproduction rate, and transparent appearance makes them highly versatile. In addition, since most compounds can easily penetrate the skin of the zebrafish, drug and toxicological screenings are able to be performed simply by adding these test compounds to their water. Zebrafish are also a widely used model system to identify endocrine disrupting chemicals (EDCs), and to perform risk assessments for human and wildlife populations.
For researchers looking at comparable nuclear receptor activity, zebrafish provide an attractive model as nearly all of the 48 human nuclear receptors are also present in the animal model. In vitro assays, such as those offered by INDIGO, allow researchers to determine the correct animal model before entering trials by providing important early indications of that compound’s potential for downstream concerns. With animal studies required by the FDA, selecting the animal model that provides the most representative human surrogate is critical to assessing a potential drug’s likelihood for unwanted effects. Cell-based assay models are crucial to help make this determination prior to entering ADMET studies.
INDIGO currently offers seven zebrafish assays:
- Zebrafish Androgen Receptor (zAR; nr3a4)
- Zebrafish Aryl Hydrocarbon Receptor (zAhR)
- Zebrafish Estrogen Receptor Alpha (zERα; nr3a1)
- Zebrafish Glucocorticoid Receptor (zGR; nr3c1)
- Zebrafish Peroxisome Proliferator-Activated Receptor Gamma (zPPARγ; nr1c3)
- Zebrafish Retinoic Acid Receptor Alpha A (zRARαa; nr1b1 isoform A)
- Zebrafish Thyroid Hormone Receptor Beta (zTRβ; nr1a2)
Kits are offered in different assay formats to accommodate researchers’ needs: 3 x 32 and 1 x 96 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications. Assay systems are all inclusive, providing reporter cells, optimized growth media, media for diluting test compounds, a positive-control agonist, luciferase detection reagent, a white assay plate, a detailed protocol, and a protocol quick guide. All kits are shipped on dry ice.
Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
The principle application of INDIGO's assay products is in the screening of test samples to quantify functional activities, either agonist or antagonist, that they may exert against the receptor. Kit products are all-inclusive assay systems that include, in addition to dFXR 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.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. 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. To initiate a Service Study, download and complete all fields of the Excel worksheet “Service Work Order" then submit the electronic file to INDIGO Customer Service.
Expansion Meets Industry Demand for Cell-Based Preclinical Models for Cancer, Chemical, and Environmental Research State College, PA (6 October 2020) – INDIGO Biosciences, the recognized industry leader in nuclear receptor research, has expanded their robust portfolio of in vitro animal model assays with three additional first-in-class zebrafish model systems. Previously available only as a live-model system, INDIGO’s
Expansion Meets Industry Demand for Pre-Clinical Models for Cancer and Environmental Research State College, PA (23 January 2020) – INDIGO Biosciences, the recognized industry leader in nuclear receptor research, has announced the addition of four zebrafish assays to its portfolio. These additions both expand INDIGO’s robust portfolio of in vitro animal model assay systems and
Disruption of Nuclear Receptor Signaling Alters Triphenyl Phosphate-Induced Cardiotoxicity in Zebrafish Embryos
ABSTRACT Triphenyl phosphate (TPHP) is an unsubstituted aryl phosphate ester used as a flame retardant and plasticizer within the United States. Using zebrafish as a model, the objectives of this study were to rely on (1) mRNA-sequencing to uncover pathways disrupted following embryonic TPHP exposure and (2) high-content screening to identify nuclear receptor ligands that