Nanoluc hdac6 fl fusion vector

HeLa cells, stabily transfected with NanoBRET plasmids NanoLuc®-HDAC6 FL Fusion Vector and NanoLuc®-HDAC10 FL Fusion Vector (Promega, Madison, WI, USA) were seeded at 20.000 cells/well in white 96 well plates. Without further incubation, tracer (0.3 µM) and drugs were added in separate steps and plates were placed in a tissue culture incubator for 2 h. For NanoBRET quantification, plates were put at room temperature for 10 minutes. Nanoglow substrate, diluted in OptiMEM without phenol red, was added and measured within 10 minutes in an OPTIMA plate reader (460 nm emission for donor and 610LP filter for acceptor). The BRET signal was calculated by the ratio of acceptor signal and donor signal.

HeLa cells, stably transfected with NanoBRET plasmids NanoLuc^®-HDAC6 FL Fusion Vector and NanoLuc^®-HDAC10 FL Fusion Vector (Promega, Madison, WI, USA) were seeded at 20,000 cells/well in white 96-well plates. Without further incubation, tracer (0.3 µM) and drugs were added in separate steps and plates were placed in a tissue culture incubator for 2 h. For NanoBRET quantification, plates were put at room temperature for 10 min. Nanoglow substrate, diluted in OptiMEM without phenol red, was added and measured within 10 min in an OPTIMA plate reader (460 nm emission for donor and 610LP filter for acceptor, BMG Labtech, Ortenberg, Germany). The BRET signal was calculated by the ratio of acceptor signal to donor signal.