Microplate washer elx50

Briefly, Corning Costar 96-Well plates were coated with NeutrAvidin at 4°C overnight. Wells were washed six times with wash buffer (water, 1 mM MnCl₂, and 1X plate wash buffer) using Microplate Washer ELx50, (Bio Tek Instruments), and then blocked with blocking buffer (HBS, 5% bovine serum albumin) for 1 hour at room temperature. Plates were coated with 1 μg/ml biotinylated cyclic V2 peptides in binding buffer (HBS, 1 mM MnCl₂) for 1 hour at room temperature. The plates were washed six times with wash buffer and HRP-conjugated anti-V2 mAb was added to wells for 1 hour at room temperature. After washing, 100 μl/well of substrate was added for 10 min to develop color at room temperature in the dark. Plates were read at OD₄₅₀ nm using an EnSpire Multimode Plate Reader (PerkinElmer).