| Assay Method Information | |
| | Competition Radioligand Binding Assay |
| Description: | Competition assays were carried out by incubation of membranes from hA1 receptors transfected to CHO cells, [3H]-DPCPX as radioligand, buffer (HEPES 20 mM (pH=7.4), 10 mM MgCl2, 100 mM NaCl, 2 units/ml adenosine deaminase), and unlabelled ligand in a total volume of 0.2 ml for 60 min at 25 C. R-PIA was used to determinate non-specific binding. Filter over Schleicher&Schuell GF/52 filters (pre-soaked 0.5% polyethylenimine) in a Brandel cell harvester. Unbound radioligand was removed with (3x 250 ul) HEPES 20 mM (pH=7.4), 100 mM NaCl and 10 mM MgCl2. Competition assays were carried out by incubation of membranes from hA2a receptors transfected to HeLa cells, [3H]ZM241385 as radioligand, buffer (50 mM Tris-HCl (pH=7.4), 10 mM MgCl2, 1 mM EDTA, 2 units/ml adenosine deaminase), and unlabelled ligand in a total volume of 0.2 ml for 30 min at 25 C. NECA was used to determinate non-specific binding. |
| Affinity data for this assay | |
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