| Assay Method Information | |
| | In Vitro Assay |
| Description: | Human SGLT-2 and SGLT-1 sequences were stably expressed in the CHO cells. The cell culture was conducted in a 96-well plate for 12 hr. The plate was washed with KRH Na+(Buffer A) or KRH-NMG (Buffer A−) buffering solution for three times, 2004/well. Then the plate was added with a buffering solution containing Buffer A or Buffer A− plus [14C]-AMG (10 μCi/mL), 100 μL/well. The cell culture was conducted at 37 °C. for 1 hr. Then, 100 μL of an ice pre-cooled buffering solution (Buffer D) was added to terminate the assay. The plate was washed for five times. Then an ice pre-cooled lytic buffering solution (100 mM NaOH solution) was added, 20 μL/well, and the centrifugation at 600 rpm was conducted for 5 mins. Then Microscint 40 solution was added, 80 μl/well, and the centrifugation at 600 rpm was conducted for 5 mins. Finally, the radioactivity of [14C]-AMG was detected with MicroBeta Trilux (purchased from PerkinElmer Co. Ltd.) according to the scintillation counting method, and the half-inhibition concentration IC50 was calculated. |
| Affinity data for this assay | |
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