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| | In Vitro Screening Assay |
| Description: | PARP-1 can catalyze the NAD-dependent addition of poly (ADP-ribose) to adjacent nuclear proteins such as histone. The activity of PARP was determined by detecting biotin coupled ADP-ribose that attaches to histone using chemiluminescence method with HT Universal Chemiluminescent PARP Assay Kit With Histone-coated Strip Wells purchased from TREVIGEN. The positive control compound was AZD-2281 purchased from SelleckChem.Method: The kit is based on ELISA. X ul sample/well, Y ul PARP enzyme/well and PARP cocktail 25 ul/well with a total volume of 50 ul/well were added into a 96-well plate coated with histone. Meanwhile, blank control (without enzyme and sample) and negative control (without sample) were set. The plate was incubated at room temperature for 60 min; washed with 1xPBS (+0.1% TritonX-100) for twice; Strep-HRP 50 ul/well was added and incubated at room temperature for 60 min; and then washed with 1xPBS (+0.1% TritonX-100) for twice. |
| Affinity data for this assay | |
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