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Thromb Haemost 1989; 61(03): 386-391
DOI: 10.1055/s-0038-1646601
Original Article
Schattauer GmbH Stuttgart

Amidolytic Detection of Prothrombin Activation Products after SDS-Gel Electrophoresis

Guido Tans
The Department of Biochemistry University of Limburg, Maastricht, The Netherlands
,
Truus Janssen-Claessen
The Department of Biochemistry University of Limburg, Maastricht, The Netherlands
,
Jan Rosing
The Department of Biochemistry University of Limburg, Maastricht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 12 December 1988

Accepted after revision 09 February 1989

Publication Date:
24 July 2018 (online)

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Summary

In this paper we report a method via which enzymatically active products formed during prothrombin activation can be detected by simple photographic means after SDS-gel electrophoresis, blotting onto a nitrocellulose membrane and visualization with the chromogenic substrate, S2238. After amidolytic detection the same nitrocellulose membrane can also be used for immunologic detection of prothrombin activation products, thus allowing a complete description of product formation during prothrombin activation.

The detection limit of the so-called “amidoblot” is approximately 3 ng thrombin per gel sample which is comparable to the sensitivity of immunoblotting.

It is further shown that the amidoblot technique can also be applied to other coagulation factors for which a suitable chromogenic substrate is available (factor XIIa, kallikrein, factor XIa, factor Xa, plasmin and activated protein C).

Keywords

Thrombin - Meizothrombin - Meizothrombin-des fragment 1 - Transblotting enzymes
 

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