The 293T cells were cotransfected with HIV-1 proviral vector pNL4-3.Luc.R−E− and VSVg-expressing vector (both obtained from the NIH AIDS Research and Reference Reagent Program). At 48–72 hours post transfection, the media containing HIV-1 virions were collected, filter-sterilized, and kept frozen. HIV-1 Luc was added to CEM T cells or PBMCs. After 48 hours, the cells were washed in PBS and lysed with Steady Lite Luciferase buffer (PerkinElmer). Light emission was analyzed in Luminoskan (PerkinElmer). To adjust for the cell number, the samples were counted with trypan blue as described in the following section.
Luminoskan
Manufactured by PerkinElmer
The Luminoskan is a microplate luminometer designed for luminescence-based assays. It measures the light emitted from samples in microplates, providing quantitative results for a variety of luminescence-based applications.
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Lab products found in correlation
2 protocols using luminoskan
1
HIV-1 Virion Production and Infection Assay
2
Measuring HIV-Luc Virus Infection
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VSVG HIV-Luc-pseudotyped HIV-1 virus was prepared as described previously [17 (link)]. CEM T cells were infected with the VSVG HIV-Luc virus and incubated overnight. The infected cells were then treated with either PP1-targeting compounds or nanoparticles and incubated overnight. Both uninfected cells and cells infected with the pseudotyped virus were used as negative and positive controls, respectively. The luciferase lysis buffer (SteadyLite, Perkin Elmer) was added to the cells and luminescence was measured using the Luminoskan (Perkin-Elmer).
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