A radioreceptor binding assay for measurement of platelet-activating factor synthesis by human neutrophils.
A new convenient method for the measurement of platelet-activating factor produced in vitro has been developed. This method involves incubation of neutrophils with stimuli, lipid extraction, purification of lipid extracts by normal phase high performance liquid chromatography (HPLC) and quantification of platelet-activating factor (PAF) by a competitive radioreceptor binding assay. The recovery of PAF from the extraction and purification procedures is 94.5 +/- 0.9%. The sensitivity of the assay is 10 pg/tube. Human neutrophils stimulated with 0, 0.25, 0.5, 1 and 2 microM A 23187 will produce ND, ND, 720, 840 and 900 pg of PAF, respectively (ND = not detectable). The values obtained for PAF produced by human neutrophils in the present assay are comparable to those obtained with the rabbit platelet aggregation assay.
Paulson
S K
SK
Department of Immunoinflammatory Diseases Research, Searle Research and Development, Skokie, IL 60077.
Nicholson
N S
NS
D001667:Binding, Competitive; D001792:Blood Platelets / Q000378:metabolism / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000096:biosynthesis*; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins; D011869:Radioligand Assay / Q000379:methods*; D011956:Receptors, Cell Surface / Q000032:analysis; D043562:Receptors, G-Protein-Coupled
D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor