產(chǎn)品概述
白細胞介素18(IL-18)是分子量為18KD的細胞因子,最早發(fā)現(xiàn)在中毒性休克中,作為共刺激因子以產(chǎn)生干擾素γ(IFN-γ)。IL-18的生物學(xué)功能與IL-12非常相似。IL-18以沒有信號肽,分子量為24KD 的前體形式被合成;有活性、成熟的IL-18必須通過切割前體產(chǎn)成。白細胞介素1β轉(zhuǎn)換酶(ICE, caspase-1)在天冬氨酸的p1位置切開IL-18前體,產(chǎn)生具有生物活性的,可以從細胞內(nèi)分泌成熟的IL-18。文獻報道,IL-18主要來源于活化的巨噬細胞,樹突狀細胞,Kupffer細胞,角質(zhì)形成細胞,腸上皮細胞,成骨細胞,腎上腺皮質(zhì)細胞和小鼠間腦等。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法??谷薎L-18單抗包被于酶標(biāo)板上,同時加入標(biāo)本、標(biāo)準品和生物素化的抗人IL-18抗體,標(biāo)本、標(biāo)準品中的IL-18會與加入于孔中的生物素化的抗人IL-18抗體及包被于酶標(biāo)板上的單抗結(jié)合,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IL-18,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, IL-18濃度與OD450值之間呈正相關(guān),可通過繪制標(biāo)準曲線求出標(biāo)本中IL-18濃度。
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