Description
Cayman’s reverse transfection reporter assays have overcome many of the disadvantages of other transfection approaches. In this method, a proprietary transfection complex containing DNA and an optimized mixture of lipids and proteins has been evenly applied and processed on the culture surface of multi-well plates. Adherent cells, supplied by the user, are applied directly to the plate and allowed to grow in the coated wells. Using this method, the uptake of the DNA complex by the cell increases dramatically compared to solution-phase transfection, enhancing both the transfection efficiency and co-transfection efficiency for multiple plasmids.Cayman’s GIP Receptor (human) Reporter Assay Kit consists of a 96-well plate coated with a transfection complex containing DNA constructs for expressing GIPR and a cAMP response element-regulated secreted alkaline phosphatase (SEAP) reporter (GIP Receptor Reverse Transfection Strip Plate). Cells grown on the transfection complex will express GIPR at the cell surface within 24 hours. Binding of agonists to GIPR initiates a signal transduction cascade through the Gαs-coupled adenylate cyclase pathway resulting in the expression of SEAP, which is secreted into the cell culture medium. Aliquots of culture medium are collected at 6-8 hours after stimulation and SEAP activity is measured following the addition of a luminescence-based alkaline phosphatase substrate provided in the kit (SEAP Substrate). The kit is easy to use and can be readily applied to high-throughput screening for therapeutic compounds regulating the activation of GIPR. A synthetic peptide agonist, tirzepatide, is included for use as a positive control. The kit provides a sufficient volume of reagents to measure SEAP activity at three time points using the three included white assay plates