SBF-1/DHT competitive assay
Flag-ARWT construct was transfected into PC3 cells and
the total protein extract was obtained and purified along with Flag tag
removal using enterokinase enzyme as reported (Skala, Goettig &
Brandstetter, 2013). SBF-1 was checked whether being binding to the
ligand binding domain of AR (AR-LBD) or not by a competitive assay using
substrate Fluormone AL Green (Thermo Fisher) according to manufacture
protocol.