Abstract: In step St1 shown in FIG. 1, a reaction system including a sample for which the content of a subject substance is to be measured, a specific-binding substance specifically binding to the subject substance, and phthalic acid or phthalate salt is constructed. The pH of the reaction system is set at less than 7. When the subject substance is an antigen, the specific-binding substance is an antibody. In reverse, when the subject substance is an antibody, the specific-binding substance is an antigen. In step St2 in FIG. 1, an optical property of the reaction system is measured. When agglutinate complexes are produced in the step St1, the reaction system becomes turbid, causing a change in scattered light intensity, transmitted light amount and the like. Therefore, by measuring the scattered light intensity, the transmitted light amount or the like, the degree of turbidity of the reaction system can be estimated.

Abstract: In step St1 shown in FIG. 1, a reaction system including a sample for which the content of a subject substance is to be measured, a specific-binding substance specifically binding to the subject substance, and phthalic acid or phthalate salt is constructed. The pH of the reaction system is set at less than 7. When the subject substance is an antigen, the specific-binding substance is an antibody. In reverse, when the subject substance is an antibody, the specific-binding substance is an antigen. In step St2 in FIG. 1, an optical property of the reaction system is measured. When agglutinate complexes are produced in the step St1, the reaction system becomes turbid, causing a change in scattered light intensity, transmitted light amount and the like. Therefore, by measuring the scattered light intensity, the transmitted light amount or the like, the degree of turbidity of the reaction system can be estimated.

Abstract: A method is provided for immunologically measuring a subject substance contained in a sample, which comprises the step of forming an antigen-antibody complex of the subject substance and a specifically binding substance capable of specifically binding to the subject substance in the presence of tricarboxylic acid or tricarboxylate and under acidic conditions. A reagent containing tricarboxylic acid or tricarboxylate for use in the method is also provided. The reagent contains a subject substance and a specifically binding substance capable of immunologically and specifically binding to the subject substance. The reagent is prepared so that the subject substance and the specifically binding substance are bound together under acidic conditions to form an antigen-antibody complex.