Reporter Assay Panel for Calcitonin and Amylin Receptors

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 Reporter Assay Panel for Calcitonin and Amylin Receptors Kit consists of a 96-well plate coated with four transfection complexes containing DNA constructs for expressing CTR (CALCR) by itself or in combination with either RAMP1, RAMP2, or RAMP3 and a cAMP response element-regulated secreted alkaline phosphatase (SEAP) reporter (CTR/AMYRs Reverse Transfection Strip Plate). Cells grown on these transfection complexes will express CTR alone (columns 1-3) or together with excess RAMP1 (columns 4-6), RAMP2 (columns 7-9), or RAMP3 (columns 10-12). These combinations form AMY₁, AMY₂, or AMY₃ receptors, respectively, on the cell surface within 24 hours. Binding of agonists initiates a signal transduction cascade through Gα_s and the adenylate cyclase pathway resulting in the expression of SEAP, which is secreted into the cell culture medium. Aliquots of culture medium are collected 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 designed primarily for evaluating specificity of ligands that activate any of the four receptors expressed on the plate. A synthetic peptide agonist, α-CGRP, 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.

Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.