预期应用
ELISA法定量测定大鼠血清、血浆、细胞培养物上清或其它相关液体中26S proteasome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26S proteasome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26S proteasome抗原、生物素化的抗大鼠26S proteasome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26S proteasome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 26S proteasome concentrations in cell culture supernates, serum, and plasma.
Introduction
The 26S proteasome is a biological macromolecule consisting of a variety of different subunits. The complex is ubiquitous in eukaryotic cells, working as natural machinery for degradation of proteins. Regarding the structure as well as the function of the 26S proteasome, the macromolecule can be subdivided into two parts. Four ring-like structures form the 20S proteasome, a cylindrically shaped molecule with a length of 15 nm and a diameter of 11 nm, enclosing three cavities. The central cavity contains the active sites of the proteasome, where peptide chains are cleaved into oligomers. Attached to the 20S core particle are one or two so-called 19S complexes, resulting in a length of 30 nm or 44 nm for the entire particle. It is assumed, that it contains the substrate binding site, recognizing proteins subjected to degradation. Additionally, a protein has to be unfolded prior to cleavage, because only the unfolded peptide chain can enter the reaction compartment of the 20S core.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for 26S proteasome has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any 26S proteasome present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for 26S proteasome 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 26S proteasome bound in the initial step. The color development is stopped and the intensity of the color is measured.
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