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Thromb Haemost 1986; 56(03): 382-386
DOI: 10.1055/s-0038-1661680
Original Article
Schattauer GmbH Stuttgart

Quantitative Immunoblotting of Plasma and Platelet Protein S

H Peter Schwarz
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California, USA
,
Mary Jo Heeb
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California, USA
,
Bernhard Lämmle
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California, USA
,
Mauro Berrettini
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California, USA
,
John H Griffin
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California, USA
› Author Affiliations
Further Information

Publication History

Received 09 June 1986

Accepted after revision 23 September 1986

Publication Date:
18 July 2018 (online)

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Summary

Protein S, the activated protein C cofactor, was measured in plasma and in platelets by a quantitative immunoblotting assay using a double antibody technique. Either whole plasma or platelet lysates were electrophoresed on sodium dodecyl sulfate polyacrylamide gels and transferred to nitrocellulose membranes by electroblotting. Based on the demonstration of proportional transfer of protein S from the gel to the nitrocellulose membrane, a reproducible and sensitive quantitative assay for protein S antigen (∼ 70,000 MW) was developed that correlated well with the Laurell rocket assay for plasma protein S. Pooled normal plasma contains 22 μg/ml protein S. Total platelet protein S antigen at ∼ 70,000 MW was determined in gel filtered platelets of ten healthy adults (mean, 163 ng per 1088 platelets). In the supernatants of thrombin-stimulated platelets, 63% of the total platelet protein S antigen was measured. Thus, quantitative immunoblotting is a useful method to detect low levels of protein S in platelets or in plasma with the advantage of giving qualitative information, i.e. apparent MW, of protein S.

Key words

Protein S - Platelets - Immunoblotting - Protein C
 

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