Kondo T, Murakami K, Isobe H, Taniguchi N, Kawakami Y
First Department of Medicine, School of Medicine, Hokkaido University, Sapporo, Japan.
Clin Chim Acta. 1990 Nov 5;191(3):169-74. doi: 10.1016/0009-8981(90)90018-n.
A simple and rapid enzyme-linked immunoassay for human erythrocyte carbonic anhydrase isozyme I was developed. The assay was found to be sensitive enough for the detection of nanogram amounts of the enzyme in incubation mixtures. The first incubation with anti-human carbonic anhydrase I IgG was carried out for 6 hours at room temperature. The second incubation of the enzyme was carried out in the presence of goat plasma C1q coupled with peroxidase for 1 h at room temperature. The enzymatic reaction was performed for 30 min using 2,2'-azino-di(3-ethyl-benzthiazoline sulfonate) as a substrate, and absorbance at 414 nm was recorded. Carbonic anhydrase I was assayed on the range of 1 to 200 ng/ml using this method. The levels of carbonic anhydrase I in K562 cells induced by erythropoietin and in other blood cells were determined. This enzyme-linked immunoassay has application for the study of developing erythroid cells.
开发了一种用于检测人红细胞碳酸酐酶同工酶I的简单快速的酶联免疫测定法。结果发现该测定法灵敏度足以检测孵育混合物中纳克量的该酶。首次与抗人碳酸酐酶I IgG孵育在室温下进行6小时。酶的第二次孵育在与过氧化物酶偶联的山羊血浆C1q存在下于室温进行1小时。使用2,2'-叠氮基二（3-乙基苯并噻唑啉磺酸盐）作为底物进行酶促反应30分钟，并记录414nm处的吸光度。使用该方法在1至200ng/ml范围内测定碳酸酐酶I。测定了促红细胞生成素诱导的K562细胞和其他血细胞中碳酸酐酶I的水平。这种酶联免疫测定法可用于研究发育中的红系细胞。
