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AssigneeKyoto Daiichi Kagaku Co., Ltd.
InventorsHarumi Uenoyama, Kyouichi Ohshiro, Atsuko Nanbu, Satoshi Fukunaga
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This invention provides a method for measuring the concentration of urinary trypsin inhibitors which is excellent in precision and reproducibility, whose operation is simple, and in which a possibility of damaging a plastic cell is eliminated. The method for measuring the concentration of urinary trypsin inhibitors comprises mixing an urine sample, a protease solution containing trypsin, and a buffer solution, adding a substrate solution to the mixture to cause the enzyme reaction, and measuring the activity of the enzyme, wherein the buffer solution is prepared so that it contains at least 0.15 .mu.mol calcium per 1 .mu.g of the trypsin but no more than 100 .mu.mol calcium per 1 ml of the urine sample in the reaction mixture, and wherein the substrate solution is prepared by dissolving the substrate in an organic solvent and diluting the mixture solution with aqueous medium, wherein at least one of an amphoteric surfactant and a nonionic surfactant is added to at least one of the organic solvent and the aqueous medium.
A method of measuring the concentration of protease inhibitor in a sample comprising mixing the sample, a protease, calcium and a substrate, and then measuring the activity of the protease, wherein the calcium content is at least 0.15 .mu.mol per 1 .mu.g of the protease but no more than 100 .mu.mol per 1 ml of the sample and wherein the substrate is used as a substrate solution containing an organic solvent, an aqueous medium and at least one amphoteric and/or nonionic surfactant.
A kit for measuring the concentration of protease inhibitor in a sample comprising a protease, a substrate and calcium, wherein the calcium content is at least 0.15 .mu.mol per 1 .mu.g of the protease but no more than 100 .mu.mol per 1 ml of the sample, and wherein the substrate is dissolved in a solution which contains an organic solvent, an aqueous medium and at least one amphoteric and/or nonionic surfactant.
A method of preparing a protease substrate solution comprising dissolving a protease substrate in an organic solvent to form a first solution and diluting the first solution with an aqueous medium, wherein at least one of an amphoteric surfactant and a nonionic surfactant is added to at least one of the organic solvent and the aqueous medium.
Dependencies are inferred from the claim text.