预期应用
ELISA法定量测定人血清、血浆或其它相关液体中γ氨基丁酸(GABA)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GABA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GABA抗原、生物素化的抗人GABA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GABA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 Gamma-aminobutyric acid,GABA concentrations in cell culture supernates, serum, and plasma.
Introduction
γ-Aminobutyricacid(GABA) has been known to function as an autocrine/paracrine signal molecule in addition to its well-known inhibitory neurotransmitter function.Studies on the developing brain and on primary brain cell cultures providede vidence for avariety of GABA functions in periods preceding the formation of synapses.
GABA is a major inhibitory neurotransmitter, the GABAergic transmission being terminated by the rapid Na+/Cl-dependent uptake through GABA transporters. It has been subdivided into neural and glial uptake systems on the basis of pharmacological properties. Recently, molecular cloning studies have identified multiple subtypes of GABA transporters (GAT1, GAT2, GAT3; and betaine GABA transporter (BGT-1). There is ~50% homology between various GABA transporter subtypes. GABA transporters are predicted to contain 12 potential transmembrane domains. The NH2 and COOH-termini are predicted to be intracellular. Two of the high affinity (Km~10 uM) rat GABA transporters (GAT2 and GAT3/GAT-B) share higher amino acid identity (68% and 65%, respectively) with the kidney betaine transporter than with GAT-1 (52%aa identity). GAT1 and GAT3 have been detected in various parts of the brain while GAT2 is found in many tissues. It appears that GAT1 and GAT3 are involved in distinct GABAergic transmission while GAT2 may be important in non-neural functions.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for GABA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any GABA present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for GABA 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 GABA bound in the initial step. The color development is stopped and the intensity of the color is measured.
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