Fig. 2 Experimental design for binding assay in microplates. Representative samples of laminin, collagen I,
collagen IV, elastin, fibronectin, BSA, and fetuin are shown in quadruplicate in the coating step (step 1). Assays
are performed in triplicate; the fourth well is used as background. Thus, after blocking, 1 μg of recombinant
protein is added in the first, second, and third columns of the plate (step 2). Anti-protein and HRP-conjugated
anti-mouse IgG are plated in all wells (step 3). A solution containing the substrate for HRP is added and
incubated for color development (step 4)
Cell Adhesion Assay for Leptospiral Proteins
181
collagen IV, elastin, fibronectin, BSA, and fetuin are shown in quadruplicate in the coating step (step 1). Assays
are performed in triplicate; the fourth well is used as background. Thus, after blocking, 1 μg of recombinant
protein is added in the first, second, and third columns of the plate (step 2). Anti-protein and HRP-conjugated
anti-mouse IgG are plated in all wells (step 3). A solution containing the substrate for HRP is added and
incubated for color development (step 4)
Cell Adhesion Assay for Leptospiral Proteins
181