It is with great pride and immense pleasure that we introduce the Creative Bioarray QualiStem® Alkaline Phosphatase Live Staining Kit which provides a fast, simple and sensitive method for AP detection without fixing the cells. The reagent is nontoxic to the cells and the fluorescent signal will be discharged from the cells by exocytosis completely within two hours after removal of the staining solution, allowing the cell to be used for follow-up studies
Details
Recombinant HEK293 cell line expressing rat PDE7A (phosphodiesterase 7A, accession number NM_031080).
N-terminal FLAG-tagged rat PDE7A has been stably expressed in a human embryonic kidney (HEK293) cell line and its expression was confirmed by Western blotting. The regulation of intracellular level of cAMP by rat PDE7A in rat PDE7A stably-expressed HEK293 cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains a luciferase gene that is under the control of the cAMP response element (CRE). When cells transiently transfected with pCRE-luc reporter were activated by forskolin, the level of cAMP was upregulated in parental HEK293 cells inducing the expression of luciferase reporter, whereas rat PDE7A-HEK293 cells showed reduction in the level of cAMP, resulting in lowered levels of luciferase expression. Inhibition of PDE7A activity by BRL 50481, a PDE7A inhibitor, restored the cAMP level, resulting in higher luciferase activity.
N-terminal FLAG-tagged rat PDE7A has been stably expressed in a human embryonic kidney (HEK293) cell line and its expression was confirmed by Western blotting. The regulation of intracellular level of cAMP by rat PDE7A in rat PDE7A stably-expressed HEK293 cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains a luciferase gene that is under the control of the cAMP response element (CRE). When cells transiently transfected with pCRE-luc reporter were activated by forskolin, the level of cAMP was upregulated in parental HEK293 cells inducing the expression of luciferase reporter, whereas rat PDE7A-HEK293 cells showed reduction in the level of cAMP, resulting in lowered levels of luciferase expression. Inhibition of PDE7A activity by BRL 50481, a PDE7A inhibitor, restored the cAMP level, resulting in higher luciferase activity.
Additional Information
| Suppliers | BPS Bioscience |
|---|---|
| Presentation | 2 vials |