| Assay Method Information | |
| | Binding at Human Serotonin Receptors Competition Binding in Human 5-HT1B Receptor Assay |
| Description: | Serotonin 5-HT1B receptor competition binding experiments were carried out in a polypropylene 96-well plate. In each well was incubated 5 μg of membranes from Hela-5-HT1B cell line prepared in the laboratory (Lot: A001/14-11-2011, protein concentration=3179 μg/ml), 1.5 nM [3H]-GR125743 (77.3 Ci/nmol, 0.1 mCi/ml, Perkin Elmer NET1172100UC) and compounds studied and standard. Non-specific binding was determined in the presence of GR55562 10 μM (TOCRIS 1054). The reaction mixture (Vt: 250 μl/well) was incubated at 25° C. for 90 min, 200 μL was transfered to GF/C 96-well plate (Millipore, Madrid, Spain) pretreated with 0.5% of PEI and treated with binding buffer (Tris-HCl 50 mM, EDTA 1 mM, MgCl2 10 mM, pH=7.4), after was filtered and washed four times with 250 μl wash buffer (Tris-HCl 50 mM, pH=7.4), before measuring in a microplate beta scintillation counter (Microbeta Trilux, PerkinElmer, Madrid, Spain). The raw radioactivity data is adjusted to a logistic regression of 4 parameters using GraphPad Prism software, from the specific junction the value of IC50 is obtained. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |