ELISA法定量测定小鼠血清、血浆或其它相关液体中磷脂酰肌醇三羟基激酶(PI3K)含量。

 

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中PI3K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PI3K、生物素化的抗小鼠PI3K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PI3K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。

This immunoassay kit allows for the specific measurement of mouse Phosphoinositide 3 kinase,PI3K concentrations in cell culture supernates, serum and plasma.

Phosphatidylinositol-3 kinase (PI3K) is a family of enzymes that phosphorylates the D3 position of phosphoinositides in membranes which can then act as a second messenger and affect many essential cellular processes such as survival, proliferation and differentiation. Class IA PI3K is composed of two subunits: a regulatory subunit, p85, and a catalytic subunit, p110. The p85 subunit is composed of several adapter domains which, upon interaction with the appropriate molecules, transmit the signal to activate p110. belongs to the  phosphoinositide 3-kinase (PI3K) family. Phosphatidylinositol 3-kinases (PI3Ks) are important constituents in insulin-like signaling pathway and involved in many critical cell functions such as proliferation, survival, motility, exocytosis, cytoskeletal rearrangements and specialized cell responses. The PI3Ks family could also be considered as oncogenes because it controls cell cycle progression, differentiation, survival, invasion and metastasis, angiogenesis, and is highly expressed in many tumors.

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for PI3K has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PI3K present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for PI3K 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 PI3K bound in the initial step. The color development is stopped and the intensity of the color is measured.