预期应用
ELISA法定量测定人血清、血浆或其它相关液体中蛋白脂质蛋白抗体(anti-PLP)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蛋白脂质蛋白抗体水平。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蛋白脂质蛋白抗体、生物素化的抗人蛋白脂质蛋白抗体抗体、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蛋白脂质蛋白抗体呈正相关。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 human proteolipid protein antibody，anti-PLP concentrations in cell culture supernates, serum, and plasma.
Introduction
Proteolipid protein (PLP) is the most abundant integral membrane protein of myelin in the central nervous system. PLP comprises 276 amino acid residues and four highly hydrophobic stretches that constitute membrane-spanning domains. A second isoform, termed DM20, is generated by alternative RNA splicing, and lacks 35 residues from an intracellular loop region. PLP acts as a strut within CNS myelin, with its extracellular loop regions defining the exact membrane-to membrane spacing in compact myelin. However, the cell biological function of PLP has been difficult to define.
PLP is associated with detergent insoluble membrane fractions (lipid rafts), but their precise role in myelin membrane formation is unclear. Using laser scanning confocal live imaging and immuno-electron microscopy, we are monitoring epitope-tagged PLP trafficking from the site of biosynthesis at the endoplasmic reticulum towards the gl ial cellular processes. This allows us to determine where myelin components are incorporated into the growing myelin sheath in vitro and in vivo.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for anti-PLP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any anti-PLP present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for anti-PLP 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 anti-PLP bound in the initial step. The color development is stopped and the intensity of the color is measured.
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