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Assay Method Information

Assay Name:  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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Last update November 1, 2007
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