ELISA experiment of Digoxin using 86100-7-RR

ELISA analysis of Digoxin was performed by coating wells of a 96-well plate with 100 µl per well of Digoxin-BSA, diluted in carbonate/bicarbonate buffer, at a concentration of 0.7 µg/mL (2hr,37℃). The plate was washed, blocked with 1% BSA blocking buffer, and incubated with 100 µl per well of Digoxin Recombinant Monoclonal Antibody (Product # 86100-7-RR) and serially diluting it with a starting concentration of 1 µg/mL (1 hr,37℃). The plate was washed and incubated with 100 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:10,000 (1 hr, 37℃). Detection was performed using an Ultra TMB Substrate (3-5 min, room temp). The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.