产品概述
γ干扰素(IFN-γ)也被称为二型干扰素,最初被认为是由促有丝分裂原激活的T淋巴细胞产生具有抗病毒活性物质。INF-γ经证实是在主体防御中起重要作用,包括免疫反应、反增生和引发炎症反应。干扰素γ主要通过T淋巴细胞、自然杀伤细胞和CD8+毒素细胞产生。另外一类型细胞包括嗜中性粒细胞、柱状细胞、角化细胞、感觉神经细胞、巨噬细胞和B细胞,这些细胞都可以有IFN-γ表达。大鼠IFN-γcDNA编码156氨基酸残基的前体蛋白和具有19个氨基酸残基的信号缩氨酸裂解产生的成熟蛋白,它包含两个潜在N端位点。天然的IFN-γ序列的三个N端(Cys-Tyr-Cys)是分离的。IFN-γ的C末端也有很高的蛋白水解活性。重组和自然产生IFN-γ以非共价连接的同型二聚体存在于溶液中。大鼠IFN-γ与小鼠IFN-γ的同源性为87%,与人IFN-γ的同源性为39%。大鼠IFN-γ对小鼠细胞有活性,对人类细胞没活性。
产品原理
本实验采用双抗体夹心ELISA法。抗大鼠IFN-γ单抗包被于酶标板上,同时加入标本、标准品和生物素化的抗大鼠IFN-γ抗体,标本、标准品中的IFN-γ会与加入于孔中的生物素化的抗大鼠IFN-γ抗体及包被于酶标板上的单抗结合,形成免疫复合物,游离的成分被洗去;加入辣根过氧化物酶标记的亲合素,亲合素与生物素特异性结合,游离的成分被洗去。加入显色底物(显色剂),若反应孔中有IFN-γ,辣根过氧化物酶会使无色的显色剂变蓝,加终止液变黄。在450nm处测OD值,IFN-γ浓度与OD450值之间呈正相关,可通过绘制标准曲线求出标本中IFN-γ浓度。
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