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Human PPARalpha
PPARα Selectivity
Service Description
Provider: INDIGO Biosciences, Inc. provides Nuclear Receptor Reporter Assay based products and services for drug discovery and toxicology. INDIGO specializes in the development of frozen, whole-cell Nuclear Receptor Assay Systems for use in small compound HTS, off-target drug interactions, and toxicology screens.
Background: Peroxisome proliferator-activated receptor alpha (PPARα), also known as NR1C1 (nuclear receptor subfamily 1, group C, member 1), is a nuclear receptor protein encoded by the PPARA gene. PPARligands, whose names derived from their effect on inducing an increase in the size and number of peroxisomes, include hypolipidemic drugs, herbicides, leukotriene antagonists, and plasticizers. Peroxisomes are subcellular organelles found in plants and animals that contain enzymes for respiration and for cholesterol and lipid metabolism. The action of peroxisome proliferators is thought to be mediated via specific receptors, called PPARs, which belong to the steroid hormone receptor superfamily. PPARs affect the expression of target genes involved in cell proliferation, cell differentiation and in immune and inflammation responses. Three closely related subtypes (alpha, beta/delta, and gamma) have been identified. Multiple alternatively spliced transcript variants have been described for PPARα gene, although the full-length nature of only two has been determined.
Service Details: This assay can be utilized to examine the agonism (activation) or antagonism (inhibition of an agonist) on the nuclear receptor. Mammalian cells are transfected with the ligand binding domain of the NR fused to a heterologous DNA binding domain as well as the response element for this DNA binding motif linked to a reporter gene. A transfection efficiency control plasmid is also included. These transfected cells are seeded in 96-well plates and then treated overnight with test compound at 37°C. Cells are then lysed and NR activation reporter activity as well as transfection efficiency reporter activity is assessed. A positive control compound (6 concentrations in triplicate) as well as vehicle control (triplicate) is included in every experiment.
For dose response curves, it is suggested that at least six concentrations of the test compounds be examined, covering a wide range of dilutions (3 log-units). In addition, we recommend that each compound/concentration be assayed in triplicate.
Deliverable: Downloadable data report containing raw and analyzed data. EC50 and peak activity will be determined using nonlinear regression. Dose-response data will be plotted for the test compounds as well as the positive control. A sample data report is available.
Price: The price of the assay includes a set-up fee, as well as a per well charge. Discounts are provided for academicians and for larger projects. Please contact us by email or phone (814) 234-1919 Ext. 303 for a price quote.
Sample Submission: The minimum sample requirement is 25 µl of a 10 mM stock solution, with the preferred sample submission of 100 mM stock solution in DMSO. A sample submission sheet is available.
Species available: Reporter assays for human, rat, mouse and cow are optimized and ready for use. Assays for other species can be custom prepared. Please visit the Custom Assay Development page for more details
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