Download PDF
Thromb Haemost 1972; 28(03): 483-488
DOI: 10.1055/s-0038-1649032
Original Article
Schattauer GmbH

Factors Affecting Factor XIII Assay by Dansyl Cadaverine Incorporation

Marion J. Sheltawy
1   University Department of Medicine, St. James’s Hospital, Leeds, England
,
Krzysztof Miloszewski
1   University Department of Medicine, St. James’s Hospital, Leeds, England
,
Monty S. Losowsky
1   University Department of Medicine, St. James’s Hospital, Leeds, England
› Author Affiliations
Further Information

Publication History

Publication Date:
29 June 2018 (online)

    Permissions and Reprints

Summary

Factors affecting the activation of factor XIII and its subsequent action in incorporating the fluorescent amine, dansyl cadaverine, into casein have been investigated.

A comparison of the activation of factor XIII at 30° C and 37° C using 5 NIH units Maws bovine thrombin showed that increasing the temperature shortened the time required for maximal activity from 4-10 minutes to 4 minutes. A sharper peak of activity was observed at the higher temperature and a more rapid fall with increasing activation time. At 30° C, 25 units Parke Davis thrombin topical gave similar results to 5 units Maws thrombin topical.

When an activation period of 10 minutes was selected, 5 units of thrombin gave the highest values of factor XIII activity.

The optimal level of calcium in the incubation medium was found to be 12 mM.

 

  • References

  • 1 Dvilansky A, Britten A. F. H, and Loewy A.Gr. 1970; Factor XIII assay by an isotope method. British Journal of Haematology 18: 399.
    CrossrefPubMedGoogle Scholar
  • 2 Loewy A. G, Dunathan K, Kriel R, and Wolfinger H. L. 1961; Fibrinase 1. Purification of substrate and enzyme. Journal of Biological Chemistry 236: 2625.
    PubMedGoogle Scholar
  • 3 Lorand L, and Konishi K. 1964; Activation of the fibrin stabilising factor of plasma by thrombin. Archives of Biochemistry 105: 58.
    CrossrefPubMedGoogle Scholar
  • 4 Lorand L, Urayama T, de Kiewiet J. W. C, and Nossel H. L. 1969; Diagnostic and genetic studies on fibrin stabilising factor with a new assay based on amine incorporation. Journal of Clinical Investigation 17: 159.
    PubMedGoogle Scholar
  • 5 Lorand L. 1969 Fibrinogen and its reactions in clotting. In: Lippincott J. B. Co. (ed.) Dynamics of Thrombus Formation and Dissolution. p. 12.
    PubMedGoogle Scholar
  • 6 Miloszewski K, Walls W. D, and Losowsky M. S. 1969; Absence of plasma transamidase activity in congenital deficiency of fibrin stabilising factor (Factor XIII). British Journal of Haematology 17: 159.
    CrossrefPubMedGoogle Scholar
  • 7 Monkhouse F. C. 1967. Antithrombin. In: Seegers W. H. (ed.) Blood Clotting Enzymology. Academic Press; New York,: 323.
    CrossrefGoogle Scholar
  • 8 Seegers W. H, Miller K. D, Andrews E. B, and Murphy R. C. 1952; Fundamental interactions and effect of storage, ether, adsorbants and blood clotting on plasma antithrombin activity. American Journal of Physiology 169: 700.
    PubMedGoogle Scholar
  • 9 Sheltawy M. J, Miloszewski K, and Losowsky M. S. 1972; Studies on the activation of human Factor XIII. Thrombosis et Diathesis Haemorrhagica 28: 473.
    PubMedGoogle Scholar