| Assay Method Information | |
| | PD-L1 Binding Assay |
| Description: | Experimental Materials:PD1:PD-L1 TR-FRET assay kit, purchased from BPS Biosciences. Nivo multimode microplate reader (PerkinElmer)Experimental Method:PD1-Eu, dye-labeled acceptor, PD-L1-biotin, and compound to be tested were all diluted using the buffer in the kit.The compound to be tested was diluted 5-fold to the 8th concentration, i.e., from 4 μM to 0.05 nM, and the DMSO concentration was 4%. The experiment was set up in duplicate wells. 5 μL of each concentration gradient of the compound to be tested was added to a microtiter plate, wherein 5 μL of a buffer containing 4% DMSO and 5 μL of PD-L1-biotin (60 nM) was added to the Max signal well, and only 5 μL of buffer was added to the Min signal well. The plate was incubated at 25° C. for 20 minutes. After the incubation, 5 μL of diluted PD1-Eu (10 nM) and 5 μL of diluted dye-labeled acceptor were added to each well. The system was reacted at 25° C. for 90 minutes. After the reaction, the multimode microplate reader was used to read the time-resolved fluorescence analysis signal. |
| Affinity data for this assay | |
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