详细信息:This assay is a Sandwich ELISA based sequentially on: 1) capture of soluble GFAP protein to the wells of a microtiter plate coated by a pre-titered amount of anti-GFAP monoclonal antibody; 2) removal of unbound material by washing; 3) binding of biotinylated anti-GFAP monoclonal antibody to
the previously captured GFAP protein; 4) removal of unbound material by washing; 5) binding of streptavidin conjugated horseradish peroxidase to the biotinylated antibodies; 6) removal of excess or unbound enzyme conjugates by washing; and 7) quantification of immobilized antibody-enzyme
complexes by monitoring horseradish peroxidase activity in the presence of the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). Enzyme activity is measured spectrophotometrically by recording absorbance readings at 450 nm. Increases in absorbency are directly proportional to the amount of
GFAP protein captured from the sample. Therefore the protein concentration of unknowns can be derived by interpolation using a standard curve generated in the same assay with reference standards of known concentrations of GFAP.
