预期应用
ELISA法定量测定人血清、血浆或其它相关液体中脂肪酸结合蛋白(FABP)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FABP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FABP、生物素化的抗人FABP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FABP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 total fatty acid-binding protein,FABP concentrations in cell culture supernates, serum, and plasma.
Introduction
Fatty acids are important for general cellular metabolism. A number of proteins have been implicated in the transport and storage of fatty acid. FABPs (fatty acid binding proteins) are a group of cytoplasmic, small mol wt (14-15 kDa) and proteins that has widespread tissue distribution. FABPs are quite abundant (3-5% of total cellular protein). The plasma kinetics of FABP (15kD) closely resemble those of myoglobin in that elevated plasma concentrations are found within 3 hour after AMI and return to normal generally within 12 to 24 hours. This makes FABP useful biochemical markers for the early assessment of exclusion of AMI.The myocardial tissue content of FABP is 5-fold lower than that of myoglobin , but the reference plasma concentration of FABP is about 15-fold lower than that of myoglobin , together suggesting a superior performance of FABP for early detection of AMI. FABP also appears a useful plasma marker for the estimation of myocardial infarct size.
At least 7 FABPs, FABP1-7, have been cloned and characterized from various tissues. FABPs can bind long-chain fatty acid, fatty-acid acyl-CoA and acyl-L-carnitine. Several different isoform of FABP have been identified and generally referred to by tissue type (liver, heart, intestine, adipocyte, kidney, brain etc; protein designated as H-FABP indicates that it is heart type). However, expression of these isoform is not exclusive and more than one isoform can be found in a given cell or tissue. Three main types of FABPs that were initially discovered in the heart (FABP-H), liver (FABP-L), and intestine (FABP-I) are not exclusive these tissues and show considerable differences at the amino acid level (~30% identity). Other FABPs recently detected in adipocyte, kidney, and brain show a high degree of sequence homology among each other and with other FABPs. FABPs are also known as mammary derived growth inhibitor (MDGI), adipocyte lipid binding protein (ALBP), Myelin protein P2 homolog, P2 adipocyte protein, 422 protein (P15). Human FABP-H or adipocyte is 132-aa protein (chromosome 2p11) Rat and mouse adipocyte-FABPs are 133 aa single polypeptide chains.Heart fatty acid-binding protein (H-FABP) is supposed to be the most sensitive biomarker of early acute myocardial infarction (AMI).
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for FABP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any FABP present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for FABP 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 FABP bound in the initial step. The color development is stopped and the intensity of the color is measured.
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