Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture ACTH from samples and standards. After washing, biotin-labeled Detection Antibody is added and incubated, followed by washing to form a "Capture Antibody-Antigen-Detection Antibody" immune-complex. Subsequently, Streptavidin-Horseradish Peroxidase (SA-HRP) is added and incubated. After incubation and washing, TMB Substrate is added for color development. If the target analyte is present in the sample, a blue color develops. Stop Solution is then added to terminate the reaction. During the assay, unbound components are washed away. The Optical Density (OD) is measured at 450 nm using a microplate reader. The intensity of the color is proportional to the ACTH concentration in the sample, and the concentration is calculated by plotting a standard curve.

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. (pg/mL)	1000	500	250	125	62.5	31.25	15.625	0
OD Value	2.708	1.531	0.792	0.422	0.229	0.131	0.086	0.039
Corrected OD Value	2.669	1.492	0.753	0.383	0.19	0.092	0.047	0

 

Sensitivity

The minimum detectable concentration (sensitivity) of Human ACTH, determined by testing samples, is 7.81pg/mL.

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