In:
Cytometry, Wiley, Vol. 48, No. 2 ( 2002-06), p. 80-86
Abstract:
Integrin αIIbβ3 mediates platelet adhesion and aggregation and plays a crucial role in thrombosis and hemostasis. αIIbβ3 is expressed in a low affinity state on resting platelets. Upon platelet activation, αIIbβ3 shifts to a high affinity conformation that efficiently binds its ligands. On human platelets, the high affinity conformation of αIIbβ3 is detected by the monoclonal antibody (mAb), PAC‐1. However, a reagent with binding specificity to high affinity mouse αIIbβ3 has not been described so far. Methods A novel rat mAb directed against mouse αIIbβ3 (JON/A) was generated and characterized. JON/A was conjugated with fluorescein isothiocyanate (JON/A FITC ) or with R‐phycoerythrin (JON/A PE ) and used for flow cytometric analysis of mouse platelets. Results Although JON/A FITC bound to resting and activated platelets, virtually no binding of the larger JON/A PE to resting platelets was detectable. However, strong binding of JON/A PE occurred on platelet activation in a dose‐dependent manner. Binding of JON/A PE required extracellular free calcium and was irreversible, thereby stabilizing the high affinity conformation of αIIbβ3. Conclusion JON/A PE is the first tool for direct assessment of integrin αIIbβ3 activation in mice. Furthermore, JON/A FITC and JON/A PE provide the first examples of fluorescent antibody derivatives with identical antigenic specificitiy that allow the discrimination between the resting and the activated state of an integrin. Cytometry 48:80–86, 2002. © 2002 Wiley‐Liss, Inc.
Type of Medium:
Online Resource
ISSN:
0196-4763
,
1097-0320
Language:
English
Publisher:
Wiley
Publication Date:
2002
detail.hit.zdb_id:
2180639-1
detail.hit.zdb_id:
1474272-X
detail.hit.zdb_id:
2180651-2
SSG:
12
