FIG. 2.

Detection of VP1 in transgenic plants by ELISA. Ninety-six-well Immulon 2 ELISA plates (Dynatech) were coated with a rabbit anti-p135-160 antiserum in carbonate buffer (pH 9.6) for 12 h at 4°C. The plates were then washed three times with PBST and blocked with 3% horse serum in PBST for 1 h at 37°C (all subsequent steps were performed with this buffer). Then, a fourfold dilution of extracts from the plants to be tested were added and incubated for 1 h at 37°C. The plates were washed, and a pool of mouse anti-p135-160 antisera was added. The plates were then washed three times and incubated for 1 h at 37°C with peroxidase-labeled rabbit anti-mouse immunoglobulin antibodies (Dakkopats). After three washes, the reaction was developed by the addition of O-phenylenediamine–H₂O₂ in citrate buffer (pH 5) and read 10 min later at 490 nm in an MR 500 Microplate Reader (Dynatech). Results are presented as rough optical density (OD) readings. Positive control is an extract of a plant transformed with pRok with the addition of p135-160 (10μg/ml).