產(chǎn)品概述
白介素-5(IL-5)是一種分泌型糖蛋白,屬于α -螺旋組細(xì)胞因子。這類蛋白主要包括 IL-3,IL-5和GM-CSF。與其他因子不同的是,IL-5 兩個(gè)鏈間是由二硫鍵共價(jià)連接形成的反平行二聚體。它主要由 CD4+ Th2細(xì)胞產(chǎn)生,活化的嗜酸性粒細(xì)胞和肥大細(xì)胞也能分泌少量的IL-5。IL-5可以促進(jìn)骨髓來(lái)源的嗜酸性粒細(xì)胞和CD34+祖細(xì)胞的產(chǎn)生和動(dòng)員。也有人認(rèn)為,由Th2細(xì)胞及肥大細(xì)胞產(chǎn)生分泌的IL-5 還可以誘導(dǎo)骨髓外嗜酸性粒細(xì)胞前體細(xì)胞成熟。人IL-5與小鼠IL-5的氨基酸序列具有70%同源性。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法。抗人IL-5單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的IL-5會(huì)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IL-5抗體,生物素化抗人IL-5抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的IL-5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(guò)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-5,辣根過(guò)氧化物酶會(huì)使無(wú)色的顯色劑變藍(lán),加終止液變黃。在450nm處測(cè)OD值,IL-5濃度與OD450值之間呈正相關(guān),可通過(guò)繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中IL-5濃度。
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