Thermo Scientific™ GeneBLAzer™ GR DA Assay Kit

Thermo Scientific™ GeneBLAzer™ GR DA Assay Kit

GeneBLAzer™GR DA (Division Arrested) cells and GR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Gluocorticoid receptor (GR) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer™UAS-bla HEK 293T cell line. GeneBLAzer™UAS-blaHEK 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)-GR (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 noapparent toxicity or change in cellular signal transduction. Both GR DA cells and GR-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of Dexamethasone. In addition, GR-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time (data available upon request). Additional testing data using alternate stimuli are also available.

Contents

• Sufficient division-arrested cells and substrate to assay one 384-well plate includes:GR HEK 293T DA cells (K1391A)LiveBLAzer™-FRET B/G Loading Kit, 70 μg