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Mouse Semaphorin 4D/CD100 ELISA Kit

时间:2016-10-28 06:39:17来源:本站 作者:admin 点击:323次


【项目简介】
Semaphorin 4D (Sema4D/CD100, previously sem J, G or C-like 2) is a 150 kDa type I transmembrane glycoprotein of the Class 4 family of transmembrane immune and nervous system semaphorins. The mouse Sema4D cDNA encodes 861 amino acids (aa) including a 23 aa signal sequence, a 710 aa extracellular domain (ECD) with sema, PSI and immunoglobulin-like domains, a 21 aa transmembrane domain and a 107 aa cytoplasmic domain. Within the ECD portion expressed (aa 24-711), mouse Sema4D shares 93%, 87%, 82% and 78% aa identity with rat, human, porcine and canine Sema4D, respectively. Sema4D is active as a homodimer that occurs via a membrane-proximal intermolecular disulfide (C707) and hydrophobic interactions involving F244 and F246. Proteolysis by metalloproteinases such as TACE/ADAM17 and MMP-14 (MT1-MMP) produces a soluble, active 120 kDa form that is also active as a dimer. Sema4D, produced by T cells and activated B and dendritic cells, acts through its low affinity receptor CD72 in the immune system. CD72 inhibits the antigen presenting cells that express it; this inhibition is relieved by Sema4D binding. 
 
【检验原理】
本试剂盒采用双抗体夹心ELISA法。酶标微孔板上预包被有抗小鼠CD100抗体,当加入标准品和待测样本时,标准品和待测样本中的小鼠CD100会和该抗体结合。洗板后,游离的其他成分被洗去,加入辣根过氧化物酶标记的的抗小鼠CD100抗体可与被酶标板捕获的小鼠CD100结合,形成“包被抗小鼠CD100抗体+小鼠CD100+抗小鼠CD100抗体-HRP”免疫复合物。当加入TMB显色底物后,复合物上连接的HRP催化底物反应,生成蓝色产物,终止反应后变为黄色。在450nm处测定OD值,小鼠CD100浓度与OD450值之间呈正比,可通过绘制标准曲线求出标本中小鼠CD100的浓度。


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