A highly sensitive chromogenic microplate assay for quantification of rat and human plasminogen.
A simple and highly sensitive chromogenic microplate assay for quantification of rat and human plasminogen in plasma samples and subcellular fractions has been developed. The assay is based on a conversion of plasminogen to plasmin, using urokinase as an activator, and a subsequent cleavage of a chromogenic plasmin substrate D-alanyl-L-cyclohexylalanyl-L-lysine-p-nitroanilide-dihydroacet ate. p-Nitroaniline being released by the cleavage is then measured at 410 nm with a microplate reader. The assay includes an acidification step to make plasminogen more readily activated to plasmin. The method is suitable for analyses of a large number of samples, measuring plasminogen in the nanogram range (0.5-50 ng/50 microliters of sample).
Department of Biochemistry, University of Olso, Norway.
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Kulseth
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Helgeland
D000818:Animals; D002863:Chromogenic Compounds; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008832:Microchemistry / Q000379:methods; D009994:Osmolar Concentration; D010958:Plasminogen / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D013997:Time Factors; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology
D002863:Chromogenic Compounds; D010958:Plasminogen; D014568:Urokinase-Type Plasminogen Activator