Tropix gal screen kit

To determine the inhibition
rate of HIV-1 by tweezers, HIV-1 NL4-3_92TH014-12 (R5) was incubated
1:1 for 10 min at 37 °C with serial dilution of tweezers, followed
by infection of TZM-bl cells in 96-well plate. Infection rates were
assessed after 48 h by detecting β-galactosidase activity in
cellular lysates using the Tropix Gal-Screen kit (Applied Biosystems)
and the Orion microplate luminometer (Berthold) for measurement. Values
represent β-galactosidase activities (relative light units per
second; RLU/s) derived from triplicate infections and normalized to
values obtained for infected cells in absence of tweezer.

To assess the
HIV-1 enhancing effect of the different amyloid fibrils, 10⁴ TZM-bl cells in 180 μL of cell culture medium were seeded
in 96-well flat-bottom plates 1 day before infection. Concentration
series of the different fibrils (0–200 μg/mL) were prepared
and then mixed with CCR5-tropic HIV-1 NL4-3 92TH014 (1 ng/mL p24 antigen).
After 10 min at room temperature, 20 μL of these mixtures were
added to the TZM-bl cells and infection rates were determined 3 days
post-infection by detecting β-galactosidase activity in cellular
lysates using the Tropix Gal-Screen kit (Applied Biosystems, Life
Technologies, Frederick, MD) and an Orion microplate luminometer (Berthold,
Bad Wildbad, Germany). All values represent reporter gene activities
(relative light units per second; RLU/s) derived from triplicate infections
minus background activities derived from uninfected cells.