An ultramicro assay for human plasma prekallikrein activity by phosphorimetry |
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Authors: | Naotaka Kuroda Hitoshi Nohta Yosuke Ohkura |
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Institution: | Faculty of Pharmaceutical Sciences, Kyushu University 62, Maidashi, Higashi-ku, Fukuoka 812 Japan |
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Abstract: | A highly sensitive method for the phosphorimetric assay of prekallikrein in human blood plasma is described. Prekallikrein is converted to kallikrein (active form) by reaction at 0°C with actin. p-Nitroaniline, formed enzymatically from H-d-prolyl-l-phenylalanyl-l-arginyl-p-nitroanilide, is extracted with ether and determined phosphorimetrically in a mixture of ether and ethanol. The method is precise and highly sensitive, requiring as little as 0.25 μl of human blood plasma. The limit of detection for p-nitroaniline formed enzymatically is 5 pmol. |
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