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The presence of a specific immunocomplex on the solid phase was detected by the action of the captured conjugate on the chromogen in the second incubation.
The biotinylated immunocomplex subsequently binds to the microparticle via interaction of biotin and streptavidin.
The amount of peroxidase retained in the immunocomplex was determined photometrically by incubating with 2,2'-azino-di-[3- ethylbenzthiazoline sulfonate] (ABTS) as a substrate for 10 min at 20[degrees]C.
The mechanism of flavonoid-induced ARF is uncertain but may be associated with hemolysis, DIC, and autoimmune and immunocomplex mechanisms.
Due to the high specificity of the interaction, little frequency change is observed unless the antigen-antibody immunocomplex is formed.
After removing the unbound antibodies, the amount of peroxidase retained in the immunocomplex was quantified by adding 2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) as the substrate, and the absorbance of the reaction mixture was measured at 405 nm using a microplate reader.