Neogen ELISA Histamine KitCortisol, or hydrocortisone, is the primary corticosteroid secreted by the adrenal cortex. Cortisol is synthesized from cholesterol and may be found in the blood as free Cortisol or bound to corticosteroid-binding globulin. The release of Cortisol is controlled by ACTH, which is produced in the anterior pituitary. Plasma Cortisol levels are highest in the morning and decrease throughout the day. Cortisol concentration in the plasma also elevates in response to stress. Cortisol has an anti-inflammatory effect and aids in carbohydrate metabolism, renal function and the promotion of glucogenesis.

Measurement of plasma Cortisol levels is useful in diagnosing conditions related to functions of the adrenal cortex, including Cushing's syndrome (hypercortisolism), Addison's disease (hypocortisolism) and adrenal tumors. Abnormal Cortisol levels may also possibly be linked to prostate cancer, depression, and schizophrenia.

This is an ELISA (Enzyme-Linked ImmunoSorbent Assay) for the quantitative analysis of Cortisol levels in biological fluid. This test kit operates based on 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 microplate. 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 650 nm. 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.

Standard Curve in EIA Buffer

Curve in EIA Buffer

Specificity (Cross-Reactivity)

Analytes Cross-Reactivity
Cortisol 100%
Prednisolone 47.4%
Cortisone 15.7%
11-Deoxycortisol 15%
Prednisone 7.83%
Corticosterone 4.81%
6β-Hydroxycortisol 1.37%
17-Hydroxyprogesterone 1.36%
Deoxycorticosterone 0.94%
Progesterone 0.06%
Bethamethasone 0.05%
Dehydroepiandrosterone 0.03%
Dexamethasone 0.03%
Beclomethasone 0.01%
d-Aldosterone 0.01%
Testosterone 0.01%
17α-Hydroxypregnenolone 0.01%
Androstenedione < 0.01%
Cholesterol < 0.01%
Estradiol < 0.01%
Estriol < 0.01%
Estrone < 0.01%
Pregnenolone < 0.01%


   80% B/B0 = 0.04 ng/mL
   50% B/B0 = 0.2 ng/mL
Assay Range: 0.04 - 10 ng/mL
Inter-Assay CV: ≤ 10%
Intra-Assay CV: ≤ 10%
Size: 96 well microplate
Assay Sample Size: 50 µL
Total Assay Incubation Time: 1.5 hours
Wavelength: 650 nm, 450 nm with acid stop
Storage Conditions: 4°C
Antibody: Polyclonal, Rabbit

Order Details

Product # Product Description
402710 Cortisol Kit, 96 well kit

Reference Materials

402710 - Cortisol, Instrucciones del kit [Inglés]

Kit inserts available on our website are for reference use only and are subject to change. It is always recommended to use the protocol included with the kit.

Calculation of Results

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