本试剂盒应用双抗体夹心酶标免疫分析法测定标本中Caspase-3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Caspase-3抗原、生物素化的抗大鼠Caspase-3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Caspase-3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 Caspase-3 concentrations in serum and plasma.
Introduction
Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires aspartic acid at the P1 position (2).
1. Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269, 30761-30764.
2. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Caspase-3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Caspase-3 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Caspase-3 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 Caspase-3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
