This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Gfap antibody was precoated onto the 96-well plate. The biotin-conjugated anti-Gfap antibody was used as the detection antibody. The standards and pilot samples were added to the wells subsequently. After incubation, unbound conjugates were removed by wash buffer. Then, a biotinylated detection antibody was added to bind with Gfap conjugated on coated antibody. After washing off unbound conjugates, HRP-Streptavidin was added. After a third washing, TMB substrates were added to visualize the HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. The concentration of Gfap in the sample was calculated by drawing a standard curve. The concentration of the target substance is proportional to the OD450 value.
