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| | Microtiter Plate Assay for Strand Transfer |
| Description: | The microtiter plate assay for stand transfer was performed with an immobilized donor substrate and a target substrate biotinylated at the 3-prime end of each DNA strand. Integrase was first assembled with immobilized LTR oligonucleotides for 30 min. Inhibitors were added after assembly and washings. For the strand transfer reaction, it was initiated with the addition of the biotinylated target substrate. Strand transfer reaction mixtures were incubated for additional 30 min, and the strand transfer products were detected by using alkaline phosphatase-conjugated avidin (Boehringer Mannheim, Indianapolis, Ind.). IC50 is the concentration of inhibitor that reduces HIV integrase activity by 50%. |
| Affinity data for this assay | |
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