| Solution Information | help | |
| Enzyme: | Envelope glycoprotein gp160 | |
| inhibitor: | BDBM61229 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Cell-cell fusion was measured following a published procedure(1), and using cell lines obtained from the NIH Research and Reference Reagents Program. TZM-bl cells expressing CD4, CCR5 and CXCR4(2), and containing an integrated reporter gene for firefly luciferase under control of HIV-1 LTR(3) were used as target cells. They were grown overnight in 96 well plates in DMEM supplemented with 10% FBS, using 25,000 cells per well. The following day, the medium was exchanged with 50 ul reduced serum medium (Gibco - Invitrogen, USA), and 1 ul of compound was added to each well, using 6 serial dilutions to obtain dose response curves. HL2/3 effector cells which produce HIV-1 HXB2 Env, Tat and Rev(4) were added in reduced serum medium, using 50,000 cells per well, to a total well volume of 100 ul. After 6 hours incubation, luciferase expression was measured using Luciferase Assay Reagent (Promega, WI, USA). Controls containing 1ul DMSO with and without HL2/3 cells were measured for each | |
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