| Assay Method Information | |
| | Determination of Affinity of Human D2L Receptors in Competitive Binding Assay Using [3H]raclopride |
| Description: | Test compounds were diluted serially in DMSO and subsequently diluted to 5% DMSO (v/w) in binding buffer (50 mM Tris, 5 mM MgCl2, 5 mM KCl, 1 mM CaCl2, 120 mM NaCl, 1 mM EDTA). A 5× concentrated compound in binding buffer (50 μL) was transferred into 96 well deep-well plates (BRAND, Wertheim, Germany). Aliquoted membrane preparations were thawed and washed once in binding buffer. In the same buffer, 10 μg protein/well was incubated with 2 nM [3H]raclopride (PerkinElmer, Waltham, MA, USA) in the presence or absence of test compound (to determine the binding inhibition of the test compound or the total binding, respectively) for 120 minutes at 25° C. in a volume of 250 μL. Non-specific binding (NSB) was determined in the presence of 10 μM haloperidol, an antagonist of the D2 receptor. |
| Affinity data for this assay | |
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