GeneBLAzer™ MR DA Assay Kit

GeneBLAzer™MR DA(Division Arrested) cells and MR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Mineralcorticoid Receptor (MR) fused to the NAbinding domain of GAL4 stably integrated in the GeneBLAzer™UAS-bla HEK 293T cell line. GeneBLAzer™UAS-bla HEK 293T cells stably express a beta-lactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS). When an agonist binds to the LBD of the GAL4 (DBD)-MR (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. Division Arrested (DA) cells are available in two configurations- an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384- well plate), and a tube of cells sufficient to analyze 10 x 384-well plates. DA cells are irreversibly division arrested using a low-dose treatment of Mitomycin-C, and have no apparent toxicity or change in cellular signal transduction. Both MR DA cells and MRUAS-bla HEK 293T cells are functionally validated for Z' and EC₅₀ concentrations of aldosterone. In addition, MR-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and substrate loading time. Additional testing data using alternate stimuli are also available.