title: In Vitro Binding of [³H]PSB-0413 to P2Y₁₂ Receptors.
abstract: The P2Y₁₂/ADP receptor plays a central role in platelet activation. Characterization of this receptor is mandatory for studying disorders associated with a P2Y₁₂ receptor defect and for evaluating P2Y₁₂ receptor agonists and antagonists. In the absence of suitable anti-P2Y₁₂ antibodies, radioligand binding assays are the only way to conduct such studies. While various radioligands were employed in the past for this purpose, none were found to be suitable for routine use. Described in this unit are protocols for quantitatively and qualitatively assessing P2Y₁₂ receptors with [³H]PSB-0413, a selective antagonist for this site. The saturation and competition assays described herein make possible the determination of P2Y₁₂ receptor density on cells, as well as the potencies and affinities of test agents at this site.
authors.affiliation: UMRS949, INSERM, Université de Strasbourg, Etablissement Français du Sang-Alsace, F-67065 Strasbourg, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Dupuis
authors.affiliation: UMRS949, INSERM, Université de Strasbourg, Etablissement Français du Sang-Alsace, F-67065 Strasbourg, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Heim
authors.affiliation: UMRS949, INSERM, Université de Strasbourg, Etablissement Français du Sang-Alsace, F-67065 Strasbourg, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Ohlmann
authors.affiliation: UMRS949, INSERM, Université de Strasbourg, Etablissement Français du Sang-Alsace, F-67065 Strasbourg, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Gachet
mesh_terms: D000249:Adenosine Monophosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D003164:Competitive Bidding / Q000379:methods; D006801:Humans; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology*; D011869:Radioligand Assay / Q000379:methods; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058925:Receptors, Purinergic P2Y12 / Q000378:metabolism*; D013872:Thionucleosides / Q000494:pharmacology*
chemical_list: C548576:P2RY12 protein, human; C511278:PSB-0413; D058921:Purinergic P2Y Receptor Antagonists; D018048:Receptors, Purinergic P2; D058925:Receptors, Purinergic P2Y12; D013872:Thionucleosides; D000249:Adenosine Monophosphate
keywords: ADP; P2Y12; binding; blood platelets; radioligand