ELISA Assays
96-well Nunc MaxisorpTM ELISA plates (Thermo Fisher)
were coated with 2µg/ml antibody in PBS at 4°C overnight. After blocking
with PBS/0.15% Casein solution for 2h at room temperature, plates were
washed 5x with PBS. Incubation of antibodies or sera was performed for
1h at RT. Afterward, plates were washed 5x with PBS/0.05% Tween. ELISAs
were developed with TMB (3,30,5,50-tetramethylbenzidine) and
H2O2 and stopped with 1mol/L sulfuric
acid after 10 minutes. OD was measured at 450 nm.