CRE Luciferase Reporter-HEK293 Cell Line

Reporter Cell Lines

Application:Monitor the GPCR-linked cAMP/PKA signaling pathway.Screen for activators or inhibitors of the GPCR-linked cAMP/PKA signaling pathway. Culture conditions:Cells should be grown at 37°C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 3 µg/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37°C-CO2  incubator.   Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence.To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly.Functional validation:A. Response of CRE  HEK293 cells to forskolin.1. Harvest CRE  HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 µl of growth medium at 5 x 10^4 c

CRE

The CRE Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the cAMP response element (CRE). The CRE   cell line is designed to monitor the cAMP/PKA signaling pathways and can be used for studying GPCR-linked cAMP/PKA signaling pathways as well as screening of agonists, antagonists or signaling inhibitors related with the cAMP/PKA signaling pathways. Functional activity of the cell line has been validated by serum stimulation assay (Figure 1).