PMID: 9426344Jan 14, 1998Paper

Detection of the three Kunitz-type single domains of membrane-bound tissue factor pathway inhibitor (TFPI) by flow cytometry

European Journal of Clinical Chemistry and Clinical Biochemistry : Journal of the Forum of European Clinical Chemistry Societies
C TiemannK Kleesiek

Abstract

Tissue factor pathway inhibitor, a natural anticoagulant in the extrinsic pathway of blood coagulation, is associated with the endothelial membrane and presumed to be released by heparin. For flow cytometric detection of membrane-bound tissue factor pathway inhibitor we synthesized polyclonal monospecific antibodies directed against each of the three Kunitz-type domains. Antisera were obtained by immunisation of rabbits with synthetic oligopeptides representing the reactive site of each domain. Kunitz-domain delta 1: 26CAFKDDGPCKAIMKR41, domain delta 2: 101EDPGICRGYITR112 and domain delta 3: 192PADRGLCRANENR204. Different cell lines (chondrosarcoma, synovial sarcoma, synovial cells, leukaemic monocytes) and endothelial cells were investigated by flow cytometric analysis using these antibodies. The three tissue factor pathway inhibitor domains were detected on the surface of all cells by the corresponding antisera. Similar results were obtained by immuno-histochemical staining. Since domain delta 3 was recognised by the appropriate antibody, it would seem that this third domain is not the membrane binding site. To investigate the cellular tissue factor pathway inhibitor release, endothelial cells were cultivated with heparin. Pr...Continue Reading

References

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Citations

Jan 17, 2013·Journal of Hematology & Oncology·Benedicte StavikNina Iversen
Dec 1, 2004·The Journal of Laboratory and Clinical Medicine·Ellen BrodinJohn-Bjarne Hansen
Aug 27, 2005·Thrombosis Research·Rafail I KushakEric F Grabowski

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