Key result
Gel affinoelectrophoresis and immunoblotting detect abnormal des-gamma-carboxylated prothrombins at a concentration of 0.1 mcg/mL.
Why the study?
A reliable and reproducible technique was needed to detect abnormal nonfunctional forms of prothrombin circulating in blood in the absence of vitamin K or presence of vitamin K antagonists.
A reliable immunoblotting technique following polyacrylamide gel affinoelectrophoresis was developed for the sensitive detection of abnormal nonfunctional prothrombin.
In the absence of vitamin K or in the presence of the vitamin K antagonists, abnormal nonfunctional forms of prothrombin circulate in the blood. A reliable and reproducible technique, derived from traditional crossed affinoimmunoelectrophoresis in presence of calcium lactate, was developed and optimized. The technique is based on nondenaturing polyacrylamide gel affinoelectrophoresis, with calcium lactate, of plasma samples, followed by immunoblotting with rabbit anti-human prothrombin serum and detection with an anti-rabbit immunoglobulin peroxidase conjugate. Depending on the plasmas, one or two bands were visualized and quantified by densitometry of the immunoblots. The technique was able to detect abnormal des-gamma-carboxylated prothrombins at concentration of 0.1 microgram/mL.
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Belle et al. (1991) studied Absence of vitamin K or presence of vitamin K antagonists. Nondenaturing polyacrylamide gel affinoelectrophoresis and immunoblotting was evaluated on Detection of abnormal des-gamma-carboxylated prothrombins. A technique based on nondenaturing polyacrylamide gel affinoelectrophoresis and immunoblotting was able to detect abnormal des-gamma-carboxylated prothrombins at a concentration of 0.1 microgram/mL.
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