预期应用
ELISA法定量测定小鼠血清、血浆、细胞培养物上清或其它相关液体中ACTH含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中ACTH水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入ACTH抗原、生物素化的抗小鼠ACTH抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的ACTH呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of mouse ACTH concentrations in cell culture supernates, serum, and plasma.
Introduction
ACTH (Adrenocorticotropic hormone) or corticotropin is a 39-amino acid peptide hormone (MW=4500) secreted by the pituitary to regulate the production of steroid hormones by the adrenal cortex. ACTH secretion from the anterior pituitary is controlled by both a classical negative feedback control mechanism and CNS-stress mediated control system. Various types of stress or pain perceived in higher levels of the brain modulate secretion of the hypothalamic neurosecretory hormone, corticotropin releasing hormone (CRH), a 41-amino acid peptide. 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. ACTH increases the synthesis and release of all adrenal sterioids, aldosterone, cortisol and adrenal androgens. It is the principal modulator of cortisol, the most important glucocorticoid in man. As the cortisol level in blood increases, release of ACTH is inhibited directly at the pituitary level. Through this same mechanism, decreasing cortisol levels lead to elevated ACTH levels. Biologically active ACTH results from enzymatic cleavage of a large precursor molecule, proopiomelanocortin (POMC). This molecule contains within its structure the amino acid sequences of ACTH, Pro-ACTH, -melanocyte stimulating hormone, lipotropin, as well as endorphin and the enkephalins.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for ACTH has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ACTH present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for ACTH is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells
and color develops in proportion to the amount of ACTH bound in the initial step. The color
development is stopped and the intensity of the color is measured.
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