Mouse/Rat ACTH Ultra Sensitive lum ELISA

Posted by admin on February 26th, 2014


SKU: AC562T-100 Category:


The CBIACTH Chemiluninescence ELISA (lumELISA) is an ultra sensitive method (Less than 1 pg/mL) intended for the quantitative determination of ACTH (Adrenocorticotropic Hormone) in Mouse/Rat plasma.

Adrenocorticotropic Hormone (ACTH) is a 39-amino acid peptide hormone (MW=4500) secreted mainly by the anterior pituitary gland. Various types of stress or pain perceived in higher levels of the brain modulate secretion of the hypothalamic neurosecretory hormone, corticotropin releasing hormone (CRH). CRH stimulates pituitary ACTH secretion. The second peptide that modulates ACTH secretion is vasopressin (AVP). AVP secretion is also stimulated by stress and acts synergistically with CRH to increase ACTH secretion in the pituitary portal circulation.

The CBIACTH Immunoassay is a two-site lumELISA (Chemiluninescence Enzyme-Linked ImmunoSorbent Assay) for the measurement of the biologically active 39 amino acid chain of ACTH. A goat polyclonal antibody to ACTH, purified by affinity chromatography, and a mouse monoclonal antibody to ACTH are specific for well-defined regions on the ACTH molecule. One antibody is prepared to bind only the C-terminal ACTH 34-39 and this antibody is biotinylated. The other antibody is prepared to bind only the mid-region and N-terminal ACTH 1-24 and this antibody is labeled with horseradish peroxidase [HRP] for detection. In this assay, calibrators, controls, or samples are simultaneously incubated with the enzyme labeled antibody and a biotin coupled antibody in a streptavidin-coated microplate well. At the end of the assay incubation, the microwell is washed to remove unbound components. Upon the addition of the luminol substrate, the enzyme activity in the enzyme-bound fraction is directly proportional to the concentration of the ACTH in the sample. A standard curve is prepared relating light unit (RLU) to the concentration of the ACTH. Concentrations of ACTH present in the controls and samples are determined directly from this curve.