Elsevier

Thrombosis Research

Volume 43, Issue 5, 1 September 1986, Pages 569-577
Thrombosis Research

Paper
Quantitation of urokinase antigen in plasma and culture media by use of an elisa

https://doi.org/10.1016/0049-3848(86)90077-0Get rights and content

Abstract

An ELISA was set up using polyvinylchloride microtiter plates coated with rabbit anti-UK IgG's and affino-purified goat anti-UK IgG's as second antibody. Detection occurred with rabbit anti-goat IgG antibodies conjugated with alkaline phosphatase. The assay is specific for urokinase (UK) with a detection limit of 100 pg/ml sample. Tissue-type plasminogen activator, up to concentrations of 100 ng/ml, does not interfere. The assay measures the antigen of the inactive zymogen pro-UK, the active enzyme UK and the UK-inhibitor complex with equal efficiency and gives the total UK antigen present, irrespective of its molecular form. Culture media of fibro blasts, endothelial- and kidney cells showed, despite the absence of active UK, antigen levels of 1.2, 23 and 65 ng/ml, respectively. In human plasma the UK concentration was found to be 3.5 ± 1.4 ng/ml (mean ± SD, n = 54). The inter- and intra-assay variations were 20% and 6%, respectively.

References (23)

  • D.J. Binnema et al.

    Characterization of type I-urokinase related plasminogen activator activities in activated plasma

    Thromb. Haemostas.

    (1985)
  • Cited by (94)

    • Plasminogen activator inhibitor type-1 deficiency does not influence the outcome of murine pneumococcal pneumonia

      2003, Blood
      Citation Excerpt :

      PAI-1 antigen and tPA antigen were measured by means of commercially available enzyme-linked immunosorbent assays (ELISAs) according to the manufacturer's recommendations: PAI-1 was measured by TintEliza PAI-1 (Biopool, Umea, Sweden); tPA, by Asserachrom tPA (Diagnostica Stago, Asnières-sur-Seine, France). The uPA antigen was measured by ELISA as described before.29 PA activity was measured by an amidolytic assay.30

    View all citing articles on Scopus
    View full text