McKimm-Breschkin J L
CSIRO Division of Biomolecular Engineering, Parkville, Australia.
J Immunol Methods. 1990 Dec 31;135(1-2):277-80. doi: 10.1016/0022-1759(90)90282-z.
A rapid, sensitive, solid-phase immunoassay, using horseradish peroxidase and 3,3',5,5'-tetramethylbenzidine, was found to be more sensitive and the colour developed was more stable compared with HRPO using 4-chloro-1-naphthol or diaminobenzidine. Assay sensitivity was equal to or greater than that obtained with the alkaline phosphatase reaction. The essential step was to incubate filters in 1% dextran sulphate for 10 min in a pH 5 (10 mM citrate-EDTA) buffer before reacting with TMB (0.05 mg/ml in the same buffer).
已发现一种使用辣根过氧化物酶和3,3',5,5'-四甲基联苯胺的快速、灵敏的固相免疫测定法，与使用4-氯-1-萘酚或二氨基联苯胺的辣根过氧化物酶法相比，该方法更灵敏，显色更稳定。测定灵敏度等于或高于碱性磷酸酶反应所获得的灵敏度。关键步骤是在与TMB（在相同缓冲液中浓度为0.05 mg/ml）反应之前，将滤膜在pH 5（10 mM柠檬酸-EDTA）缓冲液中的1%硫酸葡聚糖中孵育10分钟。
