| Assay Method Information | |
| | PARP1, PARP2 and PARP7 Enzyme Activity Inhibition Experiment |
| Description: | PARP1, PARP2 and PARP7 chemical fluorescence detection kits were all purchased from BPS Bioscience. The histone solution in the kit was diluted 5× with 1×PBS, and 25 μL of the diluted histone solution was added to a microwell plate and incubated overnight at 4° C. After the incubation was completed, the plate was washed three times with PBST (0.05% Tween-20). 25 μL of a blocking solution was added to the microwell plate and incubated at 25° C. for 90 minutes. After the incubation was completed, the plate was washed three times with PBST. 2.5 μL of compounds at different concentrations diluted in test buffer and 5 μL of 2.5× substrate mixed solution were added to the microwell plate. PARP1, PARP2 and PARP7 enzymes were diluted to 0.33, 0.23 and 5.00 ng/μL, respectively, 5 μL of the diluent was added to the microwell plate, and the reaction system was incubated at 25° C. for 60 minutes. After the incubation was completed, the plate was washed three times with PBST. Streptavidin-HRP was diluted 2000× with a blocking solution, and 25 μL of the diluent was added to the microwell plate and incubated at 25° C. for 30 minutes. After the incubation was completed, the plate was washed three times with PBST. ELISA ECL substrate A and substrate B were mixed at a ratio of 1:1 (v/v), 25 μL of the mixture was added to the microwell plate, and the chemiluminescence value was read using a BMG microplate reader. |
| Affinity data for this assay | |
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