The human platelet as a reproducible and sensitive cell for the detection and assay of platelet-activating factor.
A new procedure for the preparation of human platelets consistently sensitive to platelet-activating factor (PAF) in the low nanomolar range has been developed. Key to the success of this approach was the addition of adenosine during the isolation phase, providing an excellent recovery of stable cells, and the inclusion of ADP in the aggregation assay, providing increased sensitivity to PAF. Examination of the binding profile of tritium-labeled PAF to these platelets in the presence or absence of ADP revealed significant difference in the Kd values but not in the number of specific binding sites. Other reagents having an influence on the reactivity and stability of the human platelets, as regards its interaction with PAF, are described.
Ekholm
J
J
Department of Biochemistry, University of Texas, San Antonio 78284-7760.
Tatsumi
Y
Y
Nouchi
T
T
Hanahan
D J
DJ
D000241:Adenosine / Q000494:pharmacology; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000328:Adult; D001681:Biological Assay / Q000379:methods* / Q000706:statistics & numerical data; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D019343:Citric Acid; D005260:Female; D005340:Fibrinogen / Q000494:pharmacology; D005947:Glucose / Q000031:analogs & derivatives; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D010972:Platelet Activating Factor / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D010974:Platelet Aggregation; D015203:Reproducibility of Results; D012701:Serotonin / Q000378:metabolism
D010972:Platelet Activating Factor; C002113:acid citrate dextrose; D019343:Citric Acid; D012701:Serotonin; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D005947:Glucose; D000241:Adenosine