In the Cortisol ELISA Assay Kit, he Cortisol (Antigens) in the sample competes with the Antigensic Cortisol conjugated with horseradish peroxidase (HRP) for binding to the limited number of antibodies anti Cortisol coated on the microplate (solid phase). After incubation, the bound/free separation is performed by a simple solid-phase washing. Then, the enzyme HRP in the bound-fraction reacts with the Substrate (H2O2) and the TMB Substrate and develops a blue color that changes into yellow when the Stop Solution (H2SO4) is added. The color intensity is inversely proportional to the Cortisol concentration of in the sample. Cortisol concentration in the sample is calculated through a calibration curve. Sample Types: serum, plasma. Range: 10 - 500 ng/ml. Sensitivity: 2.44 ng/ml. Time: 1.5 hours ? Cortisol is a steroid hormone released from the adrenal cortex in response to a hormone called ACTH (produced by the pituitary gland), it is involved in the response to stress; it increases blood pressure, blood sugar levels, may cause infertility in women, and suppresses the immune system. Cortisol acts through specific intracellular receptors and has effects in numerous physiologic systems, including immune function, glucose-counter regulation, vascular tone, substrate utilization and bone metabolism. Cortisol is excreted primarily in urine in an unbound (free) form. Cortisol is bound, in plasma, from corticosteroid-binding globulin (CBG, transcotin), with high affinity, and from albumin. Only free cortisol is available to most receptors.