本试剂盒应用双抗体夹心酶标免疫分析法测定标本中KLK 6水平。用纯化的抗体包被微孔板，制成固相抗体，往包被抗体的微孔中依次加入KLK 6抗原、生物素化的抗人KLK 6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KLK 6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 Kallikrein 6,KLK 6 concentrations in cell culture supernates, serum, and plasma.
Introduction
Kallikreins (KLKs) are a subgroup of trypsin-like serine proteases and are implicated in carcinogenesis. Originally described from human urine as a hypotensive agent and later from the pancreas, kallkreins were given the Greek name for pancreas, kallikreas.
Kallikrein 6, also known as Zyme, Neurosin, PRSS9, myelencephalon-specific protease (MSP) and protease M, is a trypsin-like serine proteinase. Kallikrein 6 was originally described from the brain as an enzyme involved in degradation of amyloid plaque protein (APP) and hK6 was thought to be a beta secretase. Kallikrein 6 was shown to be elevated in the sera patients with Alzheimer’s disease and Parkinson’s disease and in animal models of multiple sclerosis. The neuronal protein alpha-synuclein was shown to be cleaved by hK6, as has APP and hK6 has been implicated in vascular morphogenesis and angiogenesis. MSP has also been shown to rapidly degrade myelin-specific protein, laminin and fibronectin. Kallikrein 6 is found in greatest abundance in the brain and neuronal tissues, with lesser levels in the kidney, thymus, spleen and lymph nodes. Human breast milk is reported to contain high levels of Kallikrein 6 and hK6 has been purified from cerebral spinal fluid (CSF). Less stringent PCR detects hK6 in many tissues and analysis of cell lines by Western blot shows that many cell lines make hK6. Endogenous inhibitors include kallistatin, protein-C inhibitor and 1-proteinase inhibitor, although Kallikrein 6 can be found complexed to a number of different proteinase inhibitors.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for Kallikrein 6 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Kallikrein 6 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for Kallikrein 6 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 Kallikrein 6 bound in the initial step. The color development is stopped and the intensity of the color is measured.
