ELISA/assay

Cortisol EIA Kit

SKT-201

96 well

Cortisol EIA Kit

Quantitative colorimetric detection of cortisol

96 well

Cell Signaling Cancer Oxidative Stress Neuroscience

(11β)-11,17,21-trihydroxypregn-4-ene-3,20-dione EIA Kit

EIA Kits

Colorimetric Assay

Competitive EIA (Enzyme Immunoassay)

Competitive

EIA kit used to quantitatively measure cortisol present in samples.

The Cortisol EIA kit is designed to quantitatively measure cortisol present in dried fecal extracts, saliva, urine, serum, plasma and tissue culture media samples. This kit measures total cortisol in extracted samples and in serum and plasma and free cortisol in saliva and urine. A cortisol standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture mouse antibodies. A cortisol-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a monoclonal antibody to cortisol to each well. After an 1 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound cortisol-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. 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.

Intra Assay Precision: Three human samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated Cortisol concentrations were: Sample 1- 1174.3 pg/mL, 6% CV Sample 2- 475.9 pg/mL, 5.6% CV Sample 3- 177.4 pg/mL, 14.7% CV Inter Assay Precision: 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: Sample 1- 1188.1 pg/mL, 7.2% CV Sample 2- 508.7 pg/mL, 6.3% CV Sample 3- 199.7 pg/mL, 10.9% CV

Species Independent

Dried Fecal Samples Saliva Urine Serum EDTA Plasma Heparin Plasma Tissue Culture Media

40 samples in duplicate

90 Minutes

17.3 pg/ml

100 - 3200 pg/ml

Cortisol, C21H30O5, (hydrocortisone, compound F) is the primary glucocorticoid produced and secreted by the adrenal cortex. 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. Immunologically, cortisol functions as an important anti-inflammatory and plays a role in hypersensitivity, immunosuppression, and disease resistance (1). In the metabolic aspect, cortisol promotes gluconeogenesis, liver glycogen deposition, and the reduction of glucose utilization (2). Production of cortisol follows an ACTH-dependent circadian rhythm, with a peak level in the morning and decreasing levels throughout the day. Most serum cortisol, all but about 4%, is bound to proteins including corticosteroid binding globulin and serum albumin (1,3). Only free cortisol is available to most receptors and it is through these receptors that physiological processes are modulated. Abnormal cortisol levels are being evaluated for correlation with a variety of different conditions, such as prostate cancer (4), depression (5), and schizophrenia (6). It is already known that abnormal levels of cortisol are involved in Cushing's Syndrome and Addision's disease (7).

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