產(chǎn)品概述
白介素-1(IL-1)是IL-1α和IL-1β 兩種蛋白的統(tǒng)稱,是由不同基因編碼的,但它們識(shí)別細(xì)胞表面相同的受體。IL-1α和IL-1β結(jié)構(gòu)相似,在氨基酸水平上有25%的同源性;兩種都是由分子量31kDa 前體裂解成17.5kDa的蛋白質(zhì)。IL-1α和IL-1β都不包含典型的疏水信號(hào)肽,但是有證據(jù)證明一些非典型路徑可以分泌這些因子。大量的IL-1α以前體形式保留在胞內(nèi)。一部分未修飾的IL-1α運(yùn)輸?shù)郊?xì)胞表面但仍與細(xì)胞相連。未修飾的膜結(jié)合IL-1α體現(xiàn)生物活性,鄰近的細(xì)胞旁分泌IL-1。 IL-1α和IL-1β與特定的受體結(jié)合后發(fā)揮其作用。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法。抗人IL-1α單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的IL-1α?xí)c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-1α抗體,生物素化抗人IL-1α抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的IL-1α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-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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