Cortisol is the most potent glucocorticoid produced and secreted by the adrenal cortex. The release of cortisol is controlled by ACTH, which is produced in the anterior pituitary. Cortisol has numerous physiological and pharmacological actions, such as carbohydrate metabolism, renal function and anti-inflammatory effect. Plasma cortisol levels are useful in diagnosis of Cushing's syndrome, Addison's disease, adrenal tumors and other conditions related to the functions of the adrenal cortex.
Principle of the Assay:
This is an ELISA for the quantitative analysis of Cortisol levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and the Cortisol in the sample for a limited number of binding sites on the antibody coated plate.
The sample or standard solution is first added to the micro-plate. Next, the diluted enzyme conjugate is added and the mixture is shaken and incubated at room temperature for one hour. During the incubation, competition for binding sites is taking place. The plate is then washed removing all the unbound material. The bound enzyme conjugate is detected by the addition of substrate, which generates an optimal color after 30 minutes.
Quantitative test results may be obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards with a microplate reader at 650nm. The extent of color development is inversely proportional to the amount of Cortisol in the sample or standard. For example, the absence of Cortisol in the sample will result in a bright blue color, whereas the presence of Cortisol will result in decreased or no color development.
C7904-20A1: ELISA Buffer, 1x30ml.
To be used to dilute enzyme conjugate and Cortisol standards.
C7904-20A2: Wash Buffer (10X), 1x20ml.
To be diluted 10x with deionized water. This is used to wash all unbound enzyme conjugate, samples and standards from the plate after the one hour incubation.
C7904-20A3: TMB Substrate, 1x20ml.
Stabilized 3,3', 5,5' Tetramethylbenzidine (TMB) plus hydrogen peroxide (H2O2) in a single bottle. It is used to develop the color in the wells after they have been washed.
C7904-20A4: Extraction Buffer (5X), 1x30ml.
To be diluted 5x with deionized water. This is used for diluting extracted and non-extracted samples.
C7904-20A5: Cortisol Enzyme Conjugate, 1x150ul.
Cortisol HRP concentrate.
C7904-20A6: Cortisol Standard, 1x100ul.Cortisol standard with a concentration of 1ug/ml.
C7904-20A7: Microtiter Strips, 1x96wells
A 96-well microtiter strips coated with rabbit anti-Cortisol in each well. The plate is ready for use as is. Do not wash.
Storage and Stability:
Store all kit components at 4 degrees C.