| Assay Method Information | |
| | In Vitro Inhibitory Activity Assay |
| Description: | The test compounds were 10-fold diluted to give a final concentration, and 10 μL of the diluted compounds was added to each well of the cell culture plate. The mixture was incubated at 37° C. for 6 h. The cells were then washed gently, the culture medium was discarded, and the cells were washed once with 200 μL of 0.1% o PBST. 50 μL of 4% PFA was added to each well, and the cells were immobilized at room temperature for 20 min, and then washed with PBS 2-3 times. 50 μL of 0.2% o Triton X-100 was added to each well, and the cells were left to stand at room temperature for 15 min and washed with 0.1% PBST 3 times. 100 uL of 3% BSA (0.6 g BSA+20 mL PBS) was added to each well, and the mixture was blocked at 37° C. for 30 min. After the blocking was completed, the liquid was removed and a primary antibody (SUMO-2/3 (181H8) Rabbit mAb, 1:400 dilution) was prepared with 100 BSA. 30 μL of the diluted primary antibody was added to each well, and the mixture was incubated at 4° C. Overnight. |
| Affinity data for this assay | |
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