Flexdrop precision reagent dispenser

We added 20 µl of bait mixture, containing 50 µM thalidomide and 0.5 µl of biotinylated HsCRBN in AlphaScreen buffer, to 384‐well AlphaPlates using a FlexDrop Precision Reagent Dispenser (PerkinElmer). We next added 0.8 µl of FLAG‐GST‐transcription factor proteins to 384‐well AlphaPlates using a NanoHead (PerkinElmer) and a Janus Workstation (PerkinElmer). After the 384‐well AlphaPlates were incubated at 26°C for 1 h, 5 µl of detection mixture containing 0.2 µg/ml anti‐DYKDDDDK mouse mAb (FUJIFILM Wako Pure Chemical), 0.08 µl of streptavidin‐coated donor beads and 0.08 µl of protein A‐coated acceptor beads (PerkinElmer) in AlphaScreen buffer were added to each well using a FlexDrop Precision Reagent Dispenser. After incubation at 26°C for 1 h, luminescent signals were detected using an EnVision plate reader (PerkinElmer).