| Assay Method Information | |
| | Binding Assay |
| Description: | Each tube in the competitive binding assay contained 100 μl membrane suspension (20 μg protein), 50 μl [125I]-AB-MECA (0.5 nM), and 50 μl of increasing concentrations of the test ligands in Tris.HCl buffer (50 mM, pH 8.0) containing 10 mM MgCl2, 1 mM EDTA. Nonspecific binding was determined using 10 μM of 5′-N-ethylcarboxamidoadenosine in the buffer. The mixtures were incubated at 25° C. for 60 min. Binding reactions were terminated by filtration through Whatman GF/B filters under reduced pressure using a MT-24 cell harvester (Brandell, Gaithersburgh, Md., USA). Filters were washed three times with 9 mL ice-cold buffer. Radioactivity was determined in a Beckman 5500B γ-counter. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |