预期应用
ELISA法定量测定小鼠血清、血浆或其它相关液体中丙酮酸脱氢酶-E1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丙酮酸脱氢酶-E1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丙酮酸脱氢酶-E1抗原、生物素化的抗小鼠丙酮酸脱氢酶-E1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丙酮酸脱氢酶-E1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 Pyruvate dehydrogenase-E1 concentrations in cell culture supernates, serum and plasma.
Introduction
Pyruvate dehydrogenase (E1) is the first component enzyme of pyruvate dehydrogenase complex (PDC). EC 1.2.4.1.Biochemical and structural data for E1s revealed a mechanism of activation of TPP cofactor by forming the conserved hydrogen bond with glutamate residue (Glu59 in human E1) and by imposing a V-conformation that brings the N4’ atom of the aminopyrimidine to the distance required for the intramolecular hydrogen bonding with the thiazolium C2 atom.
Pyruvate Dehydrogenase is a large complex containing many copies of each of three enzymes, E1, E2, and E3. The inner core of the mammalian Pyruvate Dehydrogenase complex is an icosahedral structure consisting of 60 copies of E2. At the periphery of the complex are 30 copies of E1 (itself a tetramer with subunits a2b2) and 12 copies of E3 (a homodimer), plus 12 copies of an E3 binding protein that links E3 to E2..
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Pyruvate dehydrogenase-E1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Pyruvate dehydrogenase-E1 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for Pyruvate dehydrogenase-E1 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 Pyruvate dehydrogenase-E1 bound in the initial step. The color development is stopped and the intensity of the color is measured. | 
