Assay Principle

This kit uses an indirect Competitive ELISA method. The microplate wells are pre-coated with a conjugated antigen. Cortisol in the sample competes with the pre-coated antigen for binding to the Cortisol antibody. After adding the Enzyme-labeled Secondary Antibody, a coated antigen–antibody–enzyme conjugate complex is formed. The enzyme then catalyzes the TMB substrate to produce a color reaction. The absorbance of the sample is inversely proportional to the Cortisol concentration. A standard curve is used to quantify the Cortisol content in the sample. The calculated value is multiplied by the dilution factor to obtain the actual concentration.

Typical Data

 

The following data and curve are for reference only. The experimenter must establish a standard curve based on their own experimental data.

 

Standard Conc. (ng/mL)	800	400	200	100	50	25	12.5	0
OD Value	0.174	0.235	0.337	0.524	0.835	1.223	2.012	3.061

 

Precision

Intra-assay Precision: Three samples with known high, medium, and low concentrations were assessed twenty times within the same plate.

Intra-assay Coefficient of Variation (CV%) < 10%.

 

Inter-assay Precision: Three samples with known high, medium, and low concentrations were assessed twenty times in different plates.

Inter-assay Coefficient of Variation (CV%) < 15%.

Recovery

Sample recovery rate: 80% - 120%.

Sensitivity

Based on sample testing, the minimum detectable concentration of this kit is 5.36ng/mL.

Linearity

The correlation coefficient r value of the calibrator dose-response curve is greater than or equal to 0.998.

Specificity

This kit is specific for detecting Human Cortisol and has no significant cross-reactivity with other proteins.

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