Aldosterone for accurate assessment in supernatant and serum
Cortisol, also known as hydrocortisone, is the major glucocorticoid produced and secreted by the adrenal cortex. It is involved in regulation of glucose metabolism, fat degradation, blood pressure levels, and stress inhibition. Cortisol also diffuses into cells, where its level is highly regulated by 11ß-hydroxysteroid dehydrogenase (type 1 or 2). The cortisol assay allows for rapid and accurate cortisol measurement, even in complex samples such as liver microsomes, whole cells, and animal serum. This assay can also be used to assess 11ß-HSD1 activity (native, recombinant, or microsomal).
The cortisol assay features a streamlined protocol with only two incubation steps:
- Cell stimulation by target compounds
- Cortisol detection using HTRF reagents.
This protocol requires only a single, two-hour incubation period following cell stimulation.
|Sample compatibility:||serum, whole cells, &liver microsomes|
|Detection limit:||70 pg/mL (2 fmol cortisol/well)|
|Dynamic range||0.1 to 100 ng/mL|
|Cross-reactivity:||Cortisone 0.9% |
|Z':||0.9 for 20 µl assay volumes|
Microsomal 11ß-HSD1 (0.2mg/mL) was stimulated by NADPH (200µM) and cortisone (160nM) in a 384 low-volume plate. Various concentrations of the Glycyrrhetinic acid and Carbenoxolene inhibitors were added.
A standard curve with known cortisol concentrations was run concomitantly. Plates were incubated for 2 hours at 37°C and then a cortisol-d2 conjugate and Eu3+ Cryptate labeled anti-cortisol antibodies were added.
Plates were incubated at RT for 4 hours after reagent additions and emissions read at 620 and 665 nm.
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11b-HSD1 activity measured by the HTRF cortisol assay
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In vitro inhibitory activity of 11β-HSD1assessed with the use of a HTRF...