| Assay Method Information | |
| | Binding Activity of the Compounds of the Present Disclosure for SUR1 Receptor |
| Description: | Procedures:a) 100 μL of reaction buffer was added to each well of a 96-well deep-well plate.b) 5 μL of diluted test compound (1% DMSO) was added to each well of the 96-well deep-well plate.c) 30 μL of SUR1 membrane protein and 270 μL of reaction mixture were added to each well, and the plate was shaken at 600 rpm for 5 min.d) A mixed solution of 100 μL of reaction buffer and [3H]-Glibenclamide (final concentration of 2 nM) was added to the reaction system, and the plate was shaken at 600 rpm for 5 min and incubated at 37° C. for 1 h.e) A UNIFILTER-96 GF/B plate was pretreated with 0.5% PEI, and 150 μL of 0.5% PEI was added to each well. The plate was incubated at 4° C. for 1 h.f) The UNIFILTER-96 GF/C and UNIFILTER-96 GF/B plates were washed twice with Universal Harvester, each time with 50 mL of washing solution.g) The SUR1 receptor reaction system was transferred to a UNIFILTER-96 GF/B plate with Universal Harvester, and 900 μL of washing solution was added to each well to wash the GF/B plate 4 times. The washed UNIFILTER-96 GF/B plate was dried in an oven at 55° C. for 10 min.h) 40 μL of ULTIMA GOLD scintillation solution was added to each well, and the plate was read using Microbeta. |
| Affinity data for this assay | |
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