A rapid and highly sensitive chromogenic microplate assay for quantification of rat and human prothrombin.
A rapid and highly sensitive chromogenic microplate assay for quantification of rat and human prothrombin in subcellular fractions and large series of plasma samples has been developed. The assay is based on the conversion of prothrombin to thrombin, using Echis carinatus venom as an activator, and the subsequent cleavage of a chromogenic thrombin specific substrate, D-cyclohexylglycyl-L-alanyl-L-arginine-p-nitroanilide dihydroacetate. para-Nitroaniline being released by the cleavage is then measured at 410 nm with a microplate reader. The method is suitable for analyses of a large number of samples in a short time, measuring prothrombin in the nanogram range (0.3-2.4 ng/40 microliters of sample).
Rob
J A
JA
Department of Biochemistry, University of Oslo, Norway.
Tollefsen
S
S
Helgeland
L
L
D000818:Animals; D002863:Chromogenic Compounds; D006801:Humans; D008297:Male; D011516:Prothrombin / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D013917:Thrombin / Q000187:drug effects / Q000378:metabolism; D014757:Viper Venoms / Q000378:metabolism / Q000494:pharmacology; D014859:Warfarin / Q000494:pharmacology
D002863:Chromogenic Compounds; D014757:Viper Venoms; D014859:Warfarin; D011516:Prothrombin; D013917:Thrombin