本實驗采用雙抗體夾心ELISA法??谷薉KK-1單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的DKK-1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DKK-1抗體,生物素化抗人DKK-1抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的DKK-1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(biāo)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DKK-1,辣根過氧化物酶會使無色的顯色劑變藍(lán),加終止液變黃。在450nm處測OD值, DKK-1濃度與OD450值之間呈正相關(guān),可通過繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中DKK-1濃度。
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