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Development of a competitive enzyme linked immuno sorbent assay to measure α2‐macroglobulin in irradiated rat serum

A competitive enzyme linked immunosorbent assay with antigen immobilized on the solid phase was developed to measure α2‐macroglobulin in rat serum. The cross reactivity with albumin and hemoglobin was eliminated by use of IgG fractions that were isolated after chromatography on Cibacron Blue F3‐GA Sepharose immobilized rat whole serum and hemoglobin Sepharose. Blocking materials and pH of the coating buffer had no effect on the amount of α2‐macroglobulin that binds to the plate. When the coating amount of α2‐macroglobulin was 100 ng/well, at pH 7.4, 10 mM Tris‐HCl containing 150 mM sodium chloride and the IgG amount added was 60 ng/well, then albumin and hemoglobin did not affect the assay at concentrations of 150 μg/ml or 200 μg/ml. This assay is useful for measuring the concentration of α2‐macroglobulin in normal and irradiated rat serum.