| Assay Method Information | |
| | Scintillation Proximity Assay (SPA) |
| Description: | 11beta-HSD1 microsomes isolated from HEK 293 cells over-expressing human 11beta-HSD1 were incubated with the substrate cortisone and cofactor NADPH at room temperature. The reactions were terminated with the addition of a nonspecific 11beta-HSD1 inhibitor (18beta-glycyrrhetinic acid). The product cortisol was quantified in an immuno-competition SPA wherein the [3H]-cortisol bound to anti-rabbit antibody Yitrium silicate SPA beads coated with polyclonal anti-cortisol antibody was competed by cortisol produced in the reaction, and the reaction mixture was read in a scintillation plate reader (TopCount). The amount of cortisol was determined from a cortisol standard curve. |
| Affinity data for this assay | |
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