產(chǎn)品概述
IL-1包含兩種不同基因的編碼的蛋白,IL-1α和IL-1β,但是這兩種亞型具有25%的氨基酸序列同源性,并且它們可識(shí)別相同的細(xì)胞受體。已知的分泌IL-1的細(xì)胞有成骨細(xì)胞,單核細(xì)胞,巨噬細(xì)胞,角質(zhì)形成細(xì)胞,Kupffer細(xì)胞,肝細(xì)胞,胸腺和唾液腺上皮,雪旺氏細(xì)胞,成纖維細(xì)胞和神經(jīng)膠質(zhì)細(xì)胞(少突神經(jīng)膠質(zhì),星形膠質(zhì)細(xì)胞和小膠質(zhì)細(xì)胞)。IL-1α和IL-1β都是由31KDa的前體蛋白合成。在不同的物種中,成熟的IL-1α的氨基酸序列有60%到70%的保守性,已發(fā)現(xiàn)人的IL-1可以作用于鼠的細(xì)胞系。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??勾笫驣L-1α單抗包被于酶標(biāo)板上,同時(shí)加入標(biāo)本、標(biāo)準(zhǔn)品和生物素化的抗大鼠IL-1α抗體,標(biāo)本、標(biāo)準(zhǔn)品中的IL-1α?xí)c加入于孔中的生物素化的抗大鼠IL-1α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(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-1α,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-1α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-1α濃度。
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