Enzyme electrophoresis method in analysis of active components of haemostasis system

Ostapchenko, Ludmila and Savchuk, Oleksiy and Burlova-Vasilieva, Nataliia (2011) Enzyme electrophoresis method in analysis of active components of haemostasis system. Advances in Bioscience and Biotechnology, 02 (01). pp. 20-26. ISSN 2156-8456

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Abstract

The novel modifications of substrate-containing sodium dodecyl sulfate-polyacrylamide gel electrophoresis that can be used for the detection of proteases and its activators are reported. The protease/activator samples were separated on a protein substrate-SDS-polyacrylamide gel. To detect plasminogen activators fibrinogen and Glu-plasminogen were incorporated into the SDS-PAG followed by 1 h incubation at 37?C in thrombin solution (1 NIH/ml). After electrophoresis the gel was stained according to the standard protocol. To detect fibrin-unspecific plasminogen activators from snake venom incubation in thrombin solution was substituted for 12 h incubation in 50 mM Tris-HCl (pH 7.4). To detect fibrinogen-degrading enzymes fibrinogen-containing gel was used. Activity of protease/activator was visualized in the gel as clear bands against the dark background. These new techniques offer several advantages including determination of the quantity and activity of t-PA and urokinase, however cannot be recommended for precise quantification of activators; the total procedure is quite quick and simple; method is convenient tool for detection of novel protein-protein interactions in haemostasis system; the sensitivity of the method is ≤0.01 IU per track.

Item Type: Article
Subjects: Archive Paper Guardians > Biological Science
Depositing User: Unnamed user with email support@archive.paperguardians.com
Date Deposited: 27 Mar 2023 09:03
Last Modified: 14 Mar 2024 04:44
URI: http://archives.articleproms.com/id/eprint/449

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