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Most serum cortisol, all but about 4%, is bound to proteins including corticosteroid binding globulin and serum albumin1,3. Production of cortisol follows an ACTH-dependent circadian rhythm, with a peak level in the morning and decreasing levels throughout the day. In the metabolic aspect, cortisol promotes gluconeogenesis, liver glycogen deposition, and the reduction of glucose utilization. Immunologically, cortisol functions as an important anti-inflammatory and plays a role in hypersensitivity, immunosuppression, and disease resistance. It is often referred to as the "stress hormone" as it is involved in the response to stress and it affects blood pressure, blood sugar levels, and other actions of stress adaptation. The mean and precision of the calculated Cortisol concentrations were:īackground: Cortisol, C21H30O5, (hydrocortisone, compound F) is the primary glucocorticoid produced and secreted by the adrenal cortex. Three human samples were diluted with Assay Buffer and run in duplicates in ten assays run over multiple days by four operators. The mean and precision of the calculated Cortisol concentrations were: Three human samples were diluted with Assay Buffer and run in replicates of 20 in an assay. Detectx corticosterone enzyme immunoassay kit software#The concentration of the cortisol in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader at 450 nm wavelength. The substrate reacts with the bound cortisol-peroxidase conjugate. After an hour incubation the plate is washed and substrate is added. For a further explanation go to: immunometh.pdf, page 28. Excess cortisol-peroxidase does not bind to the plates and is washed out of the well prior to the addition of substrate. The signal is generated from the cortisol-peroxidase bound to the anti-cortisol antibody which itself is bound to the goat anti-mouse IgG coated plates. As the concentration of cortisol in the sample increases, the amount of cortisol-peroxidase conjugate bound decreases causing an decrease in signal, and vice versa. The immunological reaction occurs between the limiting amount of added anti-cortisol monoclonal antibody, the cortisol antigen in the sample or standard, and the limiting amount of added cortisol-peroxidase conjugate. The binding reaction is initiated by the addition of a monoclonal antibody to cortisol. A cortisolperoxidase conjugate is added to the wells. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture mouse antibodies. ![]() A cortisol standard is provided to generate a standard curve for the assay and all samples must be read off a user-generated standard curve. Detectx corticosterone enzyme immunoassay kit free#Total cortisol is measured in extracted samples and in serum and plasma and free cortisol in saliva and urine. Please read the complete kit insert before performing this assay. The DetectX® Cortisol Immunoassay Kit is designed to quantitatively measure cortisol present in dried fecal extracts, saliva, urine, serum, plasma and tissue culture media samples. The measured concentrations were compared to the expected values based on the ratios used. Linearity was determined by taking two human urine samples diluted 1:140, one with a low diluted cortisol level of 163.9 pg/mL and one with a higher diluted level of 2,974.9 pg/mL and mixing them in the ratios given below.
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