產(chǎn)品概述
轉(zhuǎn)化生長因子(TGF)是能夠促進培養(yǎng)的成纖維細胞轉(zhuǎn)化為具有腫瘤表型的細胞的一種蛋白因子,后來人們發(fā)現(xiàn)與其腫瘤轉(zhuǎn)化作用相比,它更起著腫瘤抑制劑的作用,而且它是TGF-α和TGF-β兩種蛋白質(zhì)的混合物。這兩個蛋白質(zhì)屬于一個超家族成員,包括TGF-β1到TGF-β5骨形成蛋白質(zhì)、激活素和抑制素。人類的TGF-β1是一個 25KDa 通過二硫鍵連接在一起的沒有糖基化修飾的同源二聚體。在所有哺乳動物中幾乎有100%的序列保守性。TGF-β1生物學活性的發(fā)揮需要TGF-β1從潛態(tài)復合物中釋放出來。在體外下,可以通過分裂LAP得以實現(xiàn)(例如酸化作用)。從潛態(tài)復合物中釋放TGF存在三種最基本的活性:TGF-β1作為抑制因子調(diào)節(jié)細胞增殖;TGF-β1通過促進合成和抑制降解來提高細胞外基質(zhì)的沉積;TGF-β1通過多種機制起到免疫抑制的作用。TGF-β對特定細胞的特定作用依賴于細胞所處的環(huán)境。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法??谷薚GF-β1單抗包被于酶標板上,標本和標準品中的TGF-β1會與單抗結(jié)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TGF-β1抗體,生物素化抗人TGF-β1抗體與酶標板上結(jié)合的標準品或樣本中的TGF-β1結(jié)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(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TGF-β1,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,TGF-β1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TGF-β1濃度。
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