本试剂盒应用双抗体夹心酶标免疫分析法测定标本中D1R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D1R、生物素化的抗大鼠D1R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D1R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 Dopamine D1 receptor (D1R) concentrations in cell culture supernates, serum, and plasma.
Introduction
Dopamine is an endogenous catecholamine that influences many cellular activities, including behavior, hormone synthesis and release, blood pressure and intracellular ion transport. A family of at least 5 Dopamine Receptors (DR) genes, D1-D5, have been identified based upon the amino acid identity, pharmacological specificity and effector responses. DR have been classified into either the D1-like (D1, D1B, and D5) or D2-like (D2, D3, and D4). The two isoforms of D2R, D2 long (D2L) and short D2S), are encoded by splice variants of a single gene and differ only by the presence of an additional 29 AA in the intracellular domain 3 of the D2 long form. It may play a role in the coupling of the receptor to G-proteins. All members of this family have similar structure and contain 7 putative transmembrane domains. A given cell or tissue may express more than one DR. Specific radioligands do not exist that can differentiate between these DR.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for D1R has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any D1R present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for D1R 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 D1R bound in the initial step. The color development is stopped and the intensity of the color is measured.