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| Product Name | Cortisol EIA Kit |
|---|---|
| SKU/Catalog Number | EK7119 |
| Description | Quantitative colorimetric detection of cortisol. 96wells/kit, with removable strips. |
| Cite This Product | Cortisol EIA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7119) |
| Validated Species | All Animals |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 17.3 pg/ml *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 100 - 3200 pg/ml *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Dried Fecal Samples, EDTA Plasma, Heparin Plasma, Saliva, Serum, Tissue Culture Media, Urine *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Clear Microtitre 96 well Plate | 1 Plate |
| Cortisol Standard | 125 ?l |
| Cortisol Antibody | 3 ml |
| Cortisol Conjugate | 3 ml |
| Assay Buffer | 28 ml |
| Dissociation Reagent | 1 ml |
| Wash Buffer Concentrate | 30 ml |
| TMB Substrate | 11 ml |
| Stop Solution | 5 ml |
| Plate Cover | 1 Each |
1. Distilled or deionized water
2. Repeater pipet with disposable tips capable of dispensing 25 ?L, 50 ?L and 100 ?L.
3. Colorimetric 96 well microplate reader capable of reading optical density at 450 nm.
4. Software for converting raw relative optical density readings from the plate reader and carrying out four parameter logistic curve (4PLC) fitting. Contact your plate reader manufacturer for details.
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. In the metabolic aspect, cortisol promotes gluconeogenesis, liver glycogen deposition, and the reduction of glucose utilization. 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. 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, depression, and schizophrenia. It is already known that abnormal levels of cortisol are involved in Cushing’s Syndrome and Addision’s disease.
Click the images to enlarge.
Typical Standard Curve for the Cortisol EIA Kit (Enzyme Immunoassay)–EK7119 Assay Type: Sandwich EIA. Detection Method: Colorimetric Assay. Assay Range: 100 – 3200 pg/ml.
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. The measured concentrations were compared to the expected values based on the ratios used.
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
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.
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