本试剂盒应用双抗体夹心酶标免疫分析法测定标本中活化蛋白C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活化蛋白C、生物素化的抗人活化蛋白C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活化蛋白C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 activated protein C,APC concentrations in cell culture supernates, serum, and plasma.
Introduction
Protein C is an important player in the body’s response to inflammation, systemic sepsis and the concomitant intravascular coagulopathy. The main effect of protein C is to reduce the production of thrombin, by inactivating factors Va and VIII. As we have seen, thrombin is proinflammatory, procoagulant and antifibrinolytic. In addition, protein C inhibits the influence of tissue factor on the clotting system, reduces the production of IL-1, IL-6, and TNF-α by monocytes, and has profibrinolytic properties by inactivating PAI-1 (it inactivates the inhibitor of the activator of the agent that converts plasminogen into plasmin).
Human activated protein C is a serine protease which is derived from the two chain vitamin K dependent zymogen.Activated protein C (with protein S as a cofactor) degrades Factor Va and Factor VIIIa. Activated protein C resistance is the inability of protein C to cleave Factor Va and/or Factor VIIIa, which allows for longer duration of thrombin generation and may lead to a hypercoagulable state. This may be hereditary or acquired. The best known and most common hereditary form is Factor V Leiden. Acquired forms occur in the presence of elevated Factor VIII concentrations.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for APC has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any APC present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for APC 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 APC bound in the initial step. The color development is stopped and the intensity of the color is measured.
