| Application Notes |
To determine the anti-11D-TX antibody titer, 5 ug/ml of 11D-TX-BSA was added to an ELISA plate and incubated at 4°C overnight for antigen fixation. PBS was used as control. Unbound antigen was removed and a blocking solution was added and incubated at 37°C for an hour. The blocking solution was replaced with an appropriate dilution of antibody and incubated at 37°C for an hour. Wells were washed with PBS 3 times to remove unbound antibody, and a 1/2000 dilution of anti-mouse IgG HRP conjugated was then added as a secondary antibody. After incubation at 37°C for an hour, each well was washed with PBS 3 times to remove the unbound secondary antibody. Then TMB solution was added as substrate and incubated at 37°C for 15 minutes for color development. The reaction was stopped by the addition of stop buffer, and the absorbance was read at 450 nm with a microplate reader. |
