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、生物素化的抗大鼠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 rat Ach concentrations in Csf, serum, and plasma.
Introduction
Acetylcholine (Ach) is produced by the synthetic enzyme choline acetyltransferase which uses acetyl coenzyme A and choline as substrates for the formation of acetylcholine. Dietary choline and phosphatidylcholine serve as the sources of free choline for acetylcholine synthesis. Upon release, acetylcholine is metabolized into choline and acetate by acetylcholinesterase, and other nonspecific esterases. Acetylcholine release can be excitatory or inhibitory depending on the type of tissue and the nature of the receptor with which it interacts.
Cholinergic receptors can be divided into two types, muscarinic and nicotinic, based on the pharmacological action of various agonists and antagonists. Muscarinic receptors originally were distinguished from nicotinic receptors by the selectivity of the agonists muscarine and nicotine respectively.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Ach has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Ach present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Ach 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 Ach bound in the initial step. The color development is stopped and the intensity of the color is measured.
