Figure 1.
Vector-map, generation and purification of scuPA and scFvanti-LIBS for microbubble conjugation. A. Gene-map of scuPA in pSectag2A vector for mammalian expression. The restriction enzymes used to insert the construct are EcoRI and NotI. B. Electrophoresis with 0.8% agarose gel: pSectag2A plasmid (5137bp) after double cut restriction digest, and scuPA (924bp) after polymerase chain reaction amplification. C. Western blot analysis of scuPA after protein purification detected with an HRP-coupled anti-6x His-tag antibody, and proof of successful biotinylation of scuPA via Sortase A enzyme as detected using streptavidin HRP. D. Gene-map of scFvanti-LIBS in pAC6 vector for biotinylation. The restriction enzymes used to insert the construct are NcoI and XmaI. B. Electrophoresis with 0.8% agarose gel: pAC6 plasmid (4186bp) after double cut restriction digest, and scuPA (925bp) after polymerase chain reaction amplification. C. Western blot analysis of scFvanti-LIBS after protein purification detected with an HRP-coupled anti-6x His-tag antibody and proof of biotin on the scFvanti-LIBS as detected using streptavidin HRP.