A Highly Sensitive Chromogenic Microplate Assay for Quantification of Rat and Human Plasminogen

✍ Scribed by M.A. Kulseth; L. Helgeland

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 253 KB
• Volume: 210
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1201

✦ Synopsis

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 (\mathrm{D})-alanyl-L-cyclohexylalanyl-L-lysine(p)-nitroanilide-dihydroacetate. (p)-Nitroaniline being released by the cleavage is then measured at (410 \mathrm{~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 \mathrm{ng} / 50 \mu \mathrm{l}) of sample). C 1993 Academic Press, Inc.