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. 2007 May 30;14(8):959–968. doi: 10.1128/CVI.00123-07

FIG. 2.

FIG. 2.

GST-BKV agnoprotein purification from Sf9 cells and EIA. (A) Coomassie blue staining and detection of purified GST-agnoprotein by Western blotting by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Lane 1, cell lysate; lane 2, flowthrough; lane 3, wash; lane 4, Sepharose beads after elution; lanes 5 to 8, elution fractions 1 to 4, respectively. GST-agnoprotein was detected with monoclonal anti-GST antibody (1:5,000) and goat anti-mouse antibody conjugated to horseradish peroxidase (1:10,000). (B) A 96-well enzyme-linked immunosorbent assay plate was coated with GST-agnoprotein or GST as a control in PBS, pH 7.4 (1 pmol/well). A polyclonal rabbit anti-BKV agnoprotein serum was used as the primary antibody at a dilution of 1:20,000. Rabbit preimmune serum and PBS (pH 7.4) were used as negative controls. All assays were performed in triplicate.