| Assay Method Information | |
| | MT1/MT2 Receptor Binding Assay |
| Description: | The test tubes were placed in a reaction condition of 25° C. 10 μL of receptor membrane protein MT1 or MT2 was added to all the tubes. In non-specific binding tubes, 50 μL of a corresponding unlabeled ligand Ago at a concentration of 104 M was added to a final concentration of 10 μM, and pre-reacted for 30 minutes. In test tubes, 30 μL of test drug (MT1: a concentration of 104-10−10 M, MT2: a concentration of 104-10−11 M) was added, respectively; all tubes were added with 40 μL [3H]-Melotonin, respectively. The volume in all reaction tubes was made up to 300 μL with Tris-HCl buffer (50 mM Tris-HCl, 1 mM EDTA, 5 mM MgCl2, 0.1 mM PMSF, 0.1% NaN3, pH 7.4); the reaction was carried out at 25° C. for 1 hour; the mixture was then spotted on a type 49 glass fiber filter paper, suction filtered under a vacuum, washed 3 times with 2 mL of ice-cold Tris-HCl buffer (50 mM Tris-HCl Buffer, 1 mM EDTA, 5 mM MgCl2, 0.1% NaN3, pH7.4), and dried with suction. The filter paper was taken out and dried by baking, and then placed in a scintillation bottle. 1 mL of the scintillation liquid was added, and the radioactive intensity was measured by a liquid scintillation counter. |
| Affinity data for this assay | |
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