| Assay Method Information | |
| | Competition Binding in Human 5-HT2A Receptor |
| Description: | Serotonin 5-HT2A receptor competition binding experiments were carried out in a polypropilene 96-well plate. In each well was incubated 80 μg of membranes from CHO-5-HT2A cell line prepared in the laboratory (protein concentration=4337 μg/ml), 1 nM [3H]-Ketanserin (47.3 Ci/mmol, 1 mCi/ml, Perkin Elmer NET791250UC) and compounds studied and standard. Non-specific binding was determined in the presence of Methysergide 1 μM (Sigma M137). The reaction mixture (Vt: 250 μl/well) was incubated at 37° C. for 30 min, 200 μL was transfered to GF/B 96-well plate (Millipore, Madrid, Spain) pretreated with 0.5% of PEI and treated with binding buffer (Tris-HCl 50 mM, pH=7.4), after was filtered and washed six times with 250 μl wash buffer (Tris-HCl 50 mM, pH=6.6), before measuring in a microplate beta scintillation counter (Microbeta Trilux, PerkinElmer, Madrid, Spain). |
| Affinity data for this assay | |
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