Author: Hoffmann, Markus; González Hernández, Mariana; Berger, Elisabeth; Marzi, Andrea; Pöhlmann, Stefan
Title: The Glycoproteins of All Filovirus Species Use the Same Host Factors for Entry into Bat and Human Cells but Entry Efficiency Is Species Dependent Document date: 2016_2_22
ID: 146cwh6y_17
Snippet: For the quantification of the fLuc activity as an indicator of transduction efficiency, the cell culture supernatant was removed and the cells were washed with PBS. Next, 50 μl of 1x solution of 5x Luciferase Cell Culture Lysis Reagent (Promega) in PBS was added to each well and incubated for 30 min at room temperature, before the cell lysate was transferred to a white, opaque-walled 96well plate (Thermo Scientific). The measurement of the fLuc .....
Document: For the quantification of the fLuc activity as an indicator of transduction efficiency, the cell culture supernatant was removed and the cells were washed with PBS. Next, 50 μl of 1x solution of 5x Luciferase Cell Culture Lysis Reagent (Promega) in PBS was added to each well and incubated for 30 min at room temperature, before the cell lysate was transferred to a white, opaque-walled 96well plate (Thermo Scientific). The measurement of the fLuc activity was carried out in a microplate reader, Plate CHAMELEON (Hidex), using the MicroWin 2000 software (version 4.44, Mikrotek Laborsysteme GmbH) and fLuc substrates from the Luciferase Assay System (Promega) or Beetle-Juice (PJK) kits. Transduction efficiency, represented by fLuc activity, is either given in counts per second (cps) or displayed as x-fold change normalized against a control. All fLuc-assays were performed with quadruplicate samples.
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