| Assay Method Information | |
| | Determination of Ki Value at Human and Rat Histamin-3 Receptor |
| Description: | Histamine-3 membranes were prepared from recombinant CHO-k1 cells expressing human histamine-3 receptor or rat histamine-3 receptor. Radioligand (RS)-α-Methylhistamine [ring-1,2-3H] was purchased from American Radiolabeled chemicals (ARC) (Cat No. ART 1342). The final ligand concentration was 3 nM; non-specific determinant was Methyl histamine (100 μM) and histamine-3membrane (60 μg/well). Methyl histamine was used as a positive control. Reactions were carried out in binding buffer 50 mMTris-HCl (pH 7.4) buffer containing 5 mM MgCl2 for 60 minutes at 25° C. Incubation was stopped by rapid filtration followed by four washes of the binding mixture using 96 well harvest plate (Millipore Cat no. MSFCNXB50) pre coated with 0.5% polyethyleneimine. The plate was dried and the bound radioactivity collected on the filters was determined by scintillation counting using MicroBetaTriLux. Nonspecific binding was deducted from each data point. Radio ligand binding in the presence of non-labeled compound was expressed as a percent of the total binding and plotted against the log concentration of the compound. |
| Affinity data for this assay | |
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