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In vitro NanoLuc assay

TA Taha Azad
RS Ragunath Singaravelu
ZT Zaid Taha
TJ Taylor R. Jamieson
SB Stephen Boulton
MC Mathieu J.F. Crupi
NM Nikolas T. Martin
EB Emily E.F. Brown
JP Joanna Poutou
MG Mina Ghahremani
AP Adrian Pelin
KN Kazem Nouri
RR Reza Rezaei
CM Christopher Boyd Marshall
ME Masahiro Enomoto
RA Rozanne Arulanandam
NA Nouf Alluqmani
RS Reuben Samson
AG Anne-Claude Gingras
DC D. William Cameron
PG Peter A. Greer
CI Carolina S. Ilkow
JD Jean-Simon Diallo
JB John C. Bell
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293T cells (3 × 105 cells) were plated in 12-well plates in triplicate 24 h before transfection. Five hundred nanograms of the bioreporter constructs was transfected using PolyJet transfection reagent (SignaGen Laboratories). After 48 h, supernatant or cells lysates were collected. Cells were lysed using passive lysis buffer (Promega). NanoLuc luciferase assays were performed using one of two substrates: FMZ/ (Nano-Glo cell reagent, Promega) or native CTZ (3.33 μM final concentration; Nanolight Technologies-Prolume, Pinetop, AZ, USA). A Synergy microplate reader (BioTek, Winooski, VT, USA) was used to measure luminescence. Results are presented as RLU normalized to control. The data presented are the mean of three independent experiments.

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