预期应用
ELISA法定量测定兔血清、血浆、细胞培养物上清或其它相关液体中乙酰胆碱含量。
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Ach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Ach、生物素化的抗兔Ach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Ach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 rabbit Acetylcholine concentrations in cell culture supernates, serum, and plasma.
Introduction
Acetylcholine (Ach) is the endogenous agonist of nAChRs and is synthesized and stored in ECs and blood cells, ACh was first identified in 1914 by Henry Hallett Dale for its actions on heart tissue. It is an ester of choline and acetic acid that is the transmitter substance at many neural, or nerve, synapses and at the motor end plate of vertebrate muscles. When a nerve impulse arrives at the nerve ending, ACh, which is stored there in vesicles, is released and combines with a receptor molecule in the postsynaptic membrane or the end-plate membrane of a muscle fibre. This bonding changes the permeability of the membrane, and a change in the nature of a generator potential results. The effects of successive nerve impulses accumulate if they arrive at a sufficiently high frequency. The ACh is destroyed by an enzyme, acetylcholinesterase, and thus is effective only briefly. Inhibitors of the enzyme, however, prolong the lifetime of ACh itself. ACh affects a number of body systems including the cardiovascular system by acting as a vasodilator, by decreasing cardiac rate, and by decreasing cardiac contraction; the gastrointestinal system by such activities as increasing peristalsis in the stomach and by increasing the amplitude of digestive contractions; and the urinary tract by such actions as decreasing the capacity of the bladder and increasing the voluntary voiding pressure. It also affects the respiratory system and stimulates secretion by all glands that receive parasympathetic nerve impulses
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for Acetylcholine has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Acetylcholine present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Acetylcholine 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 Acetylcholine bound in the initial step. The color development is stopped and the intensity of the color is measured.
| 
