预期应用
ELISA法定量测定兔血清、血浆或其它相关液体中血管内皮生长因子VEGF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VEGF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VEGF抗原、生物素化的抗兔VEGF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VEGF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 Rabbit vascular endothelial growth factor (VEGF) concentrations in serum ,plasma and cell culture supernates。
Introduction
Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF) or vasculotropin , is a homodimeric 34 -42 kDa, heparin-binding glycoprotein with potent angiogenic, mitogenic and vascular permeability-enhancing activities specific for endothelial cells. The amino acid sequence of VEGF exhibits primary structural, as well as limited amino acid sequence, homology with that of the A and B chains of PDGF. VEGF is expressed by numerous rodent and tumor cells, including lung adenocarcinoma, bladder carcinoma, fibrosarcoma, HL60 promyelocytic leukemia, GS-9L glioma, and U937 lymphoma cells. In normal tissues, VEGF expression has been found in activated macrophages, keratinocytes, renal glomerular visceral epithelium and mesangial cells, hepatocytes, smooth muscle cells , Leydig cells ,embryonic fibroblasts and bronchial and choroid plexus epithelium. The expression of VEGF is upregulated by phorbol ester, TGF-a and in hypoxia. In the conditioned media of choriocarcinoma cells (JAR and JE-3), the occurrence of VEGF/PlGF heterodimers has also been observed.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for VEGF has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any VEGF present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody specific for VEGF 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 VEGF bound in the initial step. The color development is stopped and the intensity of the color is measured.
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