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| | Competitive Binding Assay |
| Description: | Competitive Binding Test of Drug to the Receptor (Rat Heart TSPO) and Radioligand (3H-PK11195): (1) tubes were placed in reaction condition of 30 C.(2) all tubes were added with 20 ug of receptor protein in order;(3) the test tubes were added in order with 20 uL of drugs in certain concentrations;(4) the non-specific binding tubes were added with 20 uL of non-tagged ligand (PK11195), the final concentration of the non-tagged ligand was 0.1 mM, pre-reacted for 30 min;(5) all tubes were added in order with 25 uL 3H-PK11195, the final concentration of the labeled ligand was 1.5 nM;(6) Tris-HCl Buffer pH7.4 was used to complement the reaction volumes of all tubes to reach 200 uL;(7) the reaction was performed under 30 C. reaction condition for 1 h;(8) Then samples were applied to 49-type glass fibre filter, vacuum suction filtered, then washed with 2 ml ice-cold Tris-HCl buffer solution (50 mM Tris-HCl buffer solution, 1 mM EDTA, 5 mM MgCl2, 1 mM PMSF, 0.1% NaN3. |
| Affinity data for this assay | |
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