Article citationsMore>>
Zhukova, A., Krasnobrysha, I. Savchuk, O. and Volkov, G. (2009) The investigation of protein-protein interactions in haemostasis system using enzyme electrophoresis method. XXII Congress of the International Society of Thrombosis and Haemostasis. Journal of Thrombosis and Haemostasis, 7, Boston, USA, 367.
has been cited by the following article:
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TITLE:
Enzyme electrophoresis method in analysis of active components of haemostasis system
AUTHORS:
Ludmila Ostapchenko, Oleksiy Savchuk, Nataliia Burlova-Vasilieva
KEYWORDS:
Substrate-Containing Electrophoresis; Enzyme Electrophoresis; Haemostasis; Proteins Activity; Proteins Identification
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.2 No.1,
February
24,
2011
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.