2.5.2 Biotinylation of the chimera ZnT8/GAD65
Biotinylation of the purified chimera was done as we have previously described (Trabucchi et al., 2019, Trabucchi et al, 2022). Two hundred eighty micrograms (280 ug) of desalted protein were incubated with excess of sulfo-NHS-biotin (Pierce Biotechnology), and free biotin was then removed.
ZnT8/GAD65-biotin was stored until use at -20 °C with the addition of 50% v/v glycerol, Tween 20 and protease inhibitors.