Fig. 4. ELISA and Western blot analysis to determine antigen binding activity of biotinylated MassM2 ScFv antibody.
Three different proteins tagged with FLAG peptide (D1.3-FLAG, GST-FLAG and DR4-FLAG) and DR4 without FLAG peptide (negative control) were coated onto 96-well plate for ELISA (A). For Western blot as shown in (B), three proteins (DR4, DR4-FLAG and GST-FLAG) were transferred onto nitrocellulose membrane. In both ELISA and Western blot, Biotinylated MassM2 ScFv antibody and streptavidin conjugated with horseradish peroxidase (HRPO) were added as primary and secondary antibody respectively. In ELISA, color development was performed using ABTS as a substrate and was measured with an 96-well plate reader at 492 nm. In Western blot, DAB was used as a substrate for final color development.
© 2022 Korean J. Microbiol.
