Fig. 4 Schematic representation of an interaction assay by ELISA. One microgram of laminin, or collagen I, or
collagen IV, or elastin, or fibronectin is adsorbed into ELISA plate in PBS buffer. BSA and fetuin are used as
control proteins. On the following day, remove the buffer, wash the plate, and block the non-specific sites of
the components using blocking buffer. After incubation for 2 h at 37
C, repeat the washing steps, and add
1 μg recombinant per well to allow interaction to the immobilized components. Bound proteins are detected by
addition of an antiserum against the recombinant protein, followed by addition of an enzyme conjugated IgG
antibody. The reaction is developed using OPD as substrate for horseradish peroxidase (HRP) that produces a
yellow-orange product detectable at 492 nm
collagen IV, or elastin, or fibronectin is adsorbed into ELISA plate in PBS buffer. BSA and fetuin are used as
control proteins. On the following day, remove the buffer, wash the plate, and block the non-specific sites of
the components using blocking buffer. After incubation for 2 h at 37
C, repeat the washing steps, and add
1 μg recombinant per well to allow interaction to the immobilized components. Bound proteins are detected by
addition of an antiserum against the recombinant protein, followed by addition of an enzyme conjugated IgG
antibody. The reaction is developed using OPD as substrate for horseradish peroxidase (HRP) that produces a
yellow-orange product detectable at 492 nm