產(chǎn)品概述
IFN-γ 的產(chǎn)生主要由活化 T 細胞產(chǎn)生,在小鼠中,由 Th1 亞群產(chǎn)生。當抗原、PHA 或 ConA 刺激后 T 細胞分泌IFN-γ ,通常與IL-2的產(chǎn)生相一致。目前認為巨噬細胞活化因子(MAF)的主要活性存在于IFN-γ中。此外,活化NK細胞也可產(chǎn)生IFN-γ。人和小鼠IFN-γ 基因分別定位于12號和10號染色體,在DNA水平上IFN-γ 基因與IFN-α /β 基因無同源性。人和小鼠 IFN-γ 在 DNA 水平上有 65%左右同源性,在氨基酸水平的同源性只有40%左右。小鼠成熟IFN-γ分子由133個氨基酸殘基組成。
人IFN-γ成熟分子是由143個氨基酸組成的糖蛋白,以同源雙體形式存在,分子量為40kDa,其生物學作用有嚴格的種屬特異性,人IFN-γ只作用于人或靈長類動物的細胞。IFN-γ 誘導單核細胞、巨噬細胞、樹突狀細胞、皮膚成纖維細胞、血管內皮細胞、星狀細胞等 MHCⅡ類抗原的表達,使其參與抗原提呈和特異性免疫的識別過程。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法??谷薎FN-γ單抗包被于酶標板上,標本和標準品中的IFN-γ會與單抗結合形成免疫復合物,游離的成分被洗去;加入生物素化的抗人IFN-γ抗體,生物素化抗人IFN-γ抗體與酶標板上結合的標準品或樣本中的IFN-γ結合而形成免疫復合物,游離的成分被洗去;加入辣根過氧化物酶標記的親合素,親合素與生物素特異性結合,游離的成分被洗去。加入顯色底物(顯色劑),若反應孔中有IFN-γ,辣根過氧化物酶會使無色的顯色劑變藍,加終止液變黃。在450nm處測OD值,IFN-γ濃度與OD450值之間呈正相關,可通過繪制標準曲線求出標本中IFN-γ濃度。
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