Development of a competitive enzyme linked immunosorbent assay to measure alpha 2-macroglobulin in irradiated rat serum.
A competitive enzyme linked immunosorbent assay with antigen immobilized on the solid phase was developed to measure alpha 2-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 alpha 2-macroglobulin that binds to the plate. When the coating amount of alpha 2-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 micrograms/ml or 200 micrograms/ml. This assay is useful for measuring the concentration of alpha 2-macroglobulin in normal and irradiated rat serum.
Kim
I G
IG
Department of Radiation Biology, Korea Atomic Energy Research Institute, Taejon, Korea.
Park
S Y
SY
D000818:Animals; D000941:Antigens / Q000302:isolation & purification; D001667:Binding, Competitive; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D005720:Gamma Rays; D007074:Immunoglobulin G / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D014916:Whole-Body Irradiation; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000276:immunology / Q000528:radiation effects*
D000941:Antigens; D007074:Immunoglobulin G; D000511:alpha-Macroglobulins