An electroblotting technique for the detection of factor VIII/von Willebrand factor multimers in plasma

British Journal of Haematology
B L Furlong, I R Peake

Abstract

A new, relatively simple technique has been developed in order to study the multimers of factor VIII/von Willebrand factor (VIII/vWF). It involves electrophoresis on SDS agarose gels and electrophoretic transfer (electroblotting) of the separated protein bands onto nitrocellulose membranes, to which they are non-covalently bound. VIII/vWF multimers are then detected by 125I-labelled antibodies to VIII/vWF, and autoradiography. Optimum electrophoretic transfer occurred at 0.5 A, for 18 h, on 0.8% agarose gels, thus enabling detection of the multimeric profile of VIII/vWF in 5 microliters of normal plasma. The multimeric profile for haemophilia A patients was identical to that for normal plasma. In plasma from patients with von Willebrand's disease (vWd), various patterns were seen, with a preponderance of smaller multimers in type II (atypical) vWd, similar to those seen in cryosupernatant. Heterogeneity within a particular type of vWd was also evident. Investigation of commercial factor VIII concentrates showed the presence of 'doublets' of VIII/vWF. Unlike other reported techniques, the rapid transfer and fixing of the protein bands to the nitrocellulose, minimizes loss of resolution, and handling of the paper is easier. It is...Continue Reading

References

Sep 1, 1978·The Journal of Clinical Investigation·R B CountsS K Elgee
Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·J C AlwineG R Stark
Sep 1, 1973·The Journal of Clinical Investigation·G A ShapiroP A McKee
Aug 1, 1972·The Journal of Clinical Investigation·S L MarchesiH R Gralnick
Nov 1, 1969·Archives of Biochemistry and Biophysics·M Steinbuch, R Audran
Jan 1, 1959·British Journal of Haematology·J G POOL, J ROBINSON
Oct 1, 1963·The Biochemical Journal·F C GREENWOODJ S GLOVER

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Citations

Jan 1, 1986·Critical Reviews in Oncology/hematology·R J HamerJ J Sixma
Jun 25, 2010·American Journal of Hematology·Marlies R Ledford-Kraemer
Dec 9, 2003·Sports Biomechanics·Pierre Gervais, John Dunn

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