Peroxisome Proliferator-Activated Receptor Alpha (PPARα; NR1C1)
|Product Family||Product Number||Product Description||Technical Manual|
|IB00111-32||Human PPARα Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB00111||Human PPARα Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB00112||Human PPARα Reporter Assay System, 1 x 384-well format assays||Technical Manual|
Peroxisome Proliferator-Activated Receptor Alpha Assay Kit
This Peroxisome Proliferator-Activated Receptor Alpha (PPARα) assay kit is an all-inclusive PPAR alpha reporter assay system that includes, in addition to PPARα 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.
PPARα 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.
INDIGO’s PPARα 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.
INDIGO's PPAR alpha assay kits are offered in different assay formats to accommodate researchers’ needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
PPARa assay kits also available in: Mouse, Rat, Cyn Monkey, Dog
Peroxisome Proliferator-Activated Receptor Alpha Assay Services
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.
PPARa Service Assays: Human, Mouse, Rat, Cyn Monkey, Dog
Peroxisome Proliferator-Activated Receptor Alpha Background
Peroxisome proliferator-activated receptor alpha (PPAR alpha), also known as NR1C1, is a nuclear receptor protein encoded by the PPARA gene. PPAR ligands, 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.
INDIGO's PPAR alpha Reporter Assay Systems utilize proprietary mammalian cells engineered to express human PPARA, commonly referred to as PPAR alpha.
The principle application of this assay product is in the screening of test samples to quantify functional activities, either agonist or antagonist, that they may exert against the human peroxisome proliferator-activated receptor alpha.
Synonyms: Peroxisome Proliferator-Activated Receptor Alpha, PPARα, PPARa, hPPARa, Human Peroxisome Proliferator-Activated Receptor Alpha, Human PPAR alpha, NR1C1
Peroxisome Proliferator-Activated Receptor Alpha Assay Data
Agonist dose-response analyses of Human PPARα. Dose-response analyses of human PPARα were performed according to the protocol provided in this Technical Manual. Reporter Cells were treated with the reference agonists GW7647 (provided), GW590735 and CP 775,146 (Tocris). Average Relative Light Units (RLU) and their respective values of Standard Deviation (SD) and Coefficient of Variation (CV) were calculated for each treatment concentration (n =4). Z’ was calculated as per Zhang, et al. (1999). Agonist treatment concentrations were Log10 transformed and respective RLU values were normalized as Fold-Activation (i.e., S/B). Data were plotted via non-linear regression and EC50 determined using GraphPad Prism software. High Z' for the reference agonist GW7647 confirms the robust performance of this human PPARα Assay.