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Effect of Anticoagulants in Colorimetric Assay for Basic Carboxypeptidases
Author(s) -
Komura Hidefumi,
Obata Kyoko,
Campbell William,
Yumoto Miho,
Shimomura Yasuyo,
Katsuya Hirotada,
Okada Noriko,
Okada Hidechika
Publication year - 2002
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2002.tb02667.x
Subject(s) - hippuric acid , carboxypeptidase , chromatography , arginine , anaphylatoxin , reagent , heparin , biochemistry , biology , chemistry , enzyme , immunology , amino acid , urine , complement system , antibody
Carboxypeptidases (CP) in plasma and sera serve as regulators of anaphylatoxins such as C3a and C5a. The activity of CP can be measured by determining hippuric acid after cleavage of the small synthetic substrate hippuryl‐ l ‐arginine. Although a colorimetric assay is convenient for determining hippuric acid generated by CP, we noticed that some anticoagulants, such as citrate, interfere with the color development of the reagents used. EDTA and heparin provide an appropriate value. EGTA used as anticoagulant also provides an appropriate value. Therefore, concentration of citrate in samples should be controlled to be constant for background subtraction.