| Assay Method Information | |
| | Time-Resolved Fluorescence (HTRF) Binding Assay |
| Description: | Inhibitors were first serially diluted in DMSO and then added to the plate wells before the addition of other reaction components. The final concentration of DMSO in the assay was 1%. The assay were carried out at 24C in the PBS buffer (pH 7.4) with 0.05% Tween-20 and 0.1% BSA. Recombinant human PD-L1 protein (19-238) with a His-tag at the C-terminus was purchased from AcroBiosystem (PDI-H5229). Recombinant human PD-1 protein (25-167) with Fc tag at the C-terminus was also purchased from AcroBiosystem (PD1-H5257). PD-L1 proteins were diluted in the assay buffer and 10ul was added to the plate well. Plates were centrifuged and proteins were preincubated with inhibitors for 40 minutes. |
| Affinity data for this assay | |
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