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Reverse fibrin autography: a method to detect and partially characterize protease inhibitors after sodium dodecyl sulfate--polyacrylamide gel electrophoresis

Academic Article
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Overview

authors

  • Erickson, L. A.
  • Lawrence, D. A.
  • Loskutoff, David J.

publication date

  • 1984

journal

  • Analytical Biochemistry  Journal

abstract

  • A new technique, reverse fibrin autography, was developed to detect protease inhibitors previously fractionated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Exogenous proteases were incorporated into fibrin-agar indicator films, eventually causing the fibrin to lyse. When an acrylamide gel containing inhibitors was placed on top of such an indicator, the positions of the inhibitors were revealed by the formation of opaque, lysis-resistant zones in the otherwise cleared fibrin film. The technique was versatile in that a variety of inhibitors were revealed, and semiquantitative since the size of the lysis-resistant zone in the indicator increased in proportion to the amount of inhibitor subjected to electrophoresis. This approach could be used not only to detect inhibitors having different protease specificities, but also to distinguish between the inhibitor activities of antibodies directed against urokinase or tissue-type plasminogen activator. Thus, reverse fibrin autography offers a convenient new approach to rapidly screen and partially characterize inhibitors present in complex biological samples.

subject areas

  • Animals
  • Antibodies
  • Cattle
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium
  • Fibrin
  • Fibrinolysis
  • Plasminogen Activators
  • Protease Inhibitors
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Identity

International Standard Serial Number (ISSN)

  • 0003-2697

Digital Object Identifier (DOI)

  • 10.1016/0003-2697(84)90113-1

PubMed ID

  • 6539574
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Additional Document Info

start page

  • 454

end page

  • 463

volume

  • 137

issue

  • 2

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