Goats were immunized with Glutathione-S-Transferase (GST) from Schistosoma japonicum. Antibody was isolated by affinity chromatography using GST immobilized on solid support and conjugated to horseradish peroxidase (HRP). Antibody concentration was determined by extinction coefficient prior to conjugation: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG. Molar enzyme/antibody protein ratio is 4:1.
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Detection of GST-tagged Protein by western blot. Samples: 0.1, 0.3, or 1.0 µg of 293 cell lysate expressing a GST-tag fusion protein. Antibodies: Affinity-purified, HRP-conjugated, goat anti-GST antibody A190-121P used for WB at 0.06 µg/ml (1:15,000). Detection: Chemiluminescence with an exposure time of 30 seconds.