Evaluation of a semiautomatic cell harvester filtration for the determination of beta-adrenoceptors in human mononuclear leukocytes.
The ligand affinity (Kd) and the number (Bmax) of beta-adrenoceptors in human mononuclear leukocytes was measured using a semiautomatic cell harvester method for the separation of bound and free ligand. The method was first evaluated in terms of contamination, nonspecific filter binding, and well-to-well reproducibility. A comparison was then made with the conventional manual filtration method. The Kd values for the binding of [125lodo]-cyanopindolol to mononuclear leukocyte membranes were independent of the method, whereas the Bmax values were systematically higher (around 30%) when determined with the cell harvester filtration method. A highly significant correlation was found between the Bmax values obtained with both methods.
Vanscheeuwijck
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P
Heymans Institute of Pharmacology, University of Ghent, Medical School, Belgium.
Fraeyman
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N
D000328:Adult; D005260:Female; D005374:Filtration / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000032:analysis*; D008024:Ligands / Q000378:metabolism; D008297:Male; D011943:Receptors, Adrenergic, beta / Q000032:analysis* / Q000378:metabolism
D008024:Ligands; D011943:Receptors, Adrenergic, beta