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- Assay Type Sandwich ELISA
- Sample Types Plasma
- Sensitivity 12.71 pg/mL
- Species Human
- Assay Duration 2 Hours
- Samples/Plate 40 in Duplicate
- Readout Colorimetric, 450 nm
- Standard Curve
The Adrenocorticotropic Hormone (ACTH) ELISA Kit quantitatively measures ACTH levels in plasma. The Adrenocorticotropic Hormone (ACTH) ELISA Kit is a sandwich ELISA with a run time of 2 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided human ACTH standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our mouse anti-human adrenocorticotropic hormone. Incubate the covered plate shaking for 1 hour at room temperature. Wash the plate and add the peroxidase-conjugated mouse anti-human ACTH monoclonal antibody. Then incubate the mixture covered at room temperature, shaking for 30 minutes. The immunological reaction occurs between the peroxidase-conjugated human ACTH antibody and the ACTH antigen in the sample or standard.
After the 30-minute incubation, wash away the excess peroxidase-conjugated human ACTH antibody and add the TMB substrate. The TMB substrate reacts with the bound peroxidase-conjugated ACTH antibody generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the ACTH concentration in the samples.
Adrenocorticotropic hormone (ACTH, adrenocorticotropin, corticotropin) is an essential component of the hypothalamic-pituitary-adrenal axis (HPA) and is produced in response to biological stress. Its principal effects are increased production and release of glucocorticoids (GCs). Stress-induced secretion of the peptide hormone Corticotropin Releasing Hormone (CRH) stimulates pituitary ACTH secretion. Circulating ACTH binds to melanocortin receptors on the surface of adrenal zona cells, inducing the synthesis and release of all adrenal steroids, aldosterone, GCs, and adrenal androgens. ACTH is also the principal modulator of cortisol and corticosterone. In addition to the stress response, ACTH synthesis correlates with the circadian rhythm in many organisms. Measuring plasma ACTH is helpful in the differential diagnosis of pituitary Cushing’s disease, Addison’s disease, adrenal tumors, adrenal hyperplasia, and ectopic ACTH syndrome.