產(chǎn)品概述
白細(xì)胞介素 4(IL-4)是由活化的T淋巴細(xì)胞、肥大細(xì)胞、嗜堿細(xì)胞產(chǎn)生的多功能性細(xì)胞因子。IL-4對一系列細(xì)胞起多種免疫調(diào)節(jié)活性。它在誘導(dǎo)IgE在B淋巴細(xì)胞的同型轉(zhuǎn)換中起重要的調(diào)控作用。它在前體T輔助細(xì)胞到Th2亞類介導(dǎo)體液免疫和調(diào)節(jié)抗體產(chǎn)生同型變換中起重要的調(diào)節(jié)作用。除此之外,IL-4在體內(nèi)外都表明有抗腫瘤活性。小鼠cDNA序列編碼140個氨基酸殘基的前體蛋白質(zhì),其中包括20個氨基酸殘基的信號肽,當(dāng)信號肽切割后就成為成熟蛋白質(zhì)。在氨基酸序列水平,成熟小鼠的IL-4與人的IL-4有50%的同源性,但這兩個蛋白質(zhì)的生物活性沒有種屬的交叉反應(yīng)。小鼠的IL-4與小鼠的IL-13有30%的氨基酸序列同源性,并且這兩個分子呈現(xiàn)出重疊的生物活性。IL-4基因定位到小鼠的11號染色體上,接近IL-3,IL-5,IL-13和GM-CSF(粒細(xì)胞-巨噬細(xì)胞集落刺激因子)的基因。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法??剐∈驣L-4單抗包被于酶標(biāo)板上,加入標(biāo)準(zhǔn)品和待測標(biāo)本,標(biāo)準(zhǔn)品和標(biāo)本中的IL-4會與包被于酶標(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;加入生物素化的抗小鼠IL-4抗體和辣根過氧化物酶標(biāo)記的親合素,抗小鼠IL-4抗體與被捕獲的標(biāo)準(zhǔn)品及待測標(biāo)本結(jié)合,生物素與親和素特異性結(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-4,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,IL-4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IL-4濃度。
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