產(chǎn)品概述
TNF-α,也稱為 TNFSFIA,是腫瘤壞死因子家族的成員,在炎癥反應(yīng)與細胞凋亡中發(fā)揮重要作用。初始合成時是26kD分子量的233個氨基酸組成II型跨膜蛋白質(zhì),包含30個氨基酸組成的胞內(nèi)區(qū), 26 個氨基酸組成的跨膜區(qū),以及177個氨基酸組成的胞外區(qū)。TNF-α胞內(nèi)通過非共價的連接形成跨膜的同源二聚體。通過 TNF-α 轉(zhuǎn)換酶(TACE)的作用,從跨膜區(qū)C端釋放出一個由157個氨基酸組成的可溶 TNF-α(s TNF-α 。目前,已知表達TNF-α的人類細胞有:B細胞,結(jié)腸柱狀上皮細胞,NK細胞, CD3+CD56+肝自然T細胞,巨噬細胞,單核細胞,樹突狀細胞,CD4+ CD8+ T細胞,肥大細胞,嗜中性粒細胞,角化細胞,血漿細胞以及脂肪細胞等。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法??谷薚NF-α單抗包被于酶標板上,標本和標準品中的TNF-α會與單抗結(jié)合形成免疫復(fù)合物,游離的成分被洗去;加入生物素化的抗人TNF-α抗體,生物素化抗人TNF-α抗體與酶標板上結(jié)合的標準品或樣本中的TNF-α結(jié)合而形成免疫復(fù)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(jié)合,游離的成分被洗去。加入顯色底物(顯色劑),若反應(yīng)孔中有TNF-α,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,TNF-α濃度與OD450值之間呈正相關(guān),可通過繪制標準曲線求出標本中TNF-α濃度。
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