產(chǎn)品概述
白細(xì)胞介素-4是一種多效性的細(xì)胞因子,它在不同種類的細(xì)胞類型中有多種免疫反應(yīng)調(diào)節(jié)活性。IL-4是一種用于調(diào)節(jié)免疫球蛋白同型轉(zhuǎn)換,特別是IgG1和IgE的B細(xì)胞激活或活化因子。它抑制IFN-γ產(chǎn)生CD4+T細(xì)胞并且調(diào)控T細(xì)胞前體向Th2亞群的分化,調(diào)節(jié)過(guò)敏和體液免疫反應(yīng)。TNF-α,IL-4協(xié)同誘導(dǎo)VCAM-1在血管內(nèi)皮細(xì)胞和平滑肌細(xì)胞的表達(dá),致使嗜曙紅細(xì)胞和淋巴細(xì)胞選擇性誘發(fā)這個(gè)位置的發(fā)炎。IL-4下游調(diào)控炎癥介質(zhì),如IL-1、TNF-α、和PGE2在單核細(xì)胞中產(chǎn)生。IL-4同樣被證實(shí)在體內(nèi)或體外有抗腫瘤活性作用。已經(jīng)證明能分泌IL-4的細(xì)胞包括嗜堿細(xì)胞,CD8+T細(xì)胞,記憶CD4+ 細(xì)胞,前體TH2細(xì)胞,柱狀細(xì)胞,嗜曙紅細(xì)胞,病毒活性的樹(shù)枝狀細(xì)胞。鼠的IL-4cDNA序列預(yù)測(cè)是一個(gè)147個(gè)氨基酸殘基的前體蛋白包括一個(gè)24個(gè)氨基酸殘基的信號(hào)肽序列,通過(guò)剪切產(chǎn)生了123個(gè)氨基酸殘基的成熟蛋白,包括4個(gè)潛在的N端的糖基化位點(diǎn)和7個(gè)半胱氨酸殘基,這7個(gè)中的6個(gè)參與形成三個(gè)分子內(nèi)的有活性的二硫鍵連接。成熟大鼠的IL-4氨基酸序列與小鼠和人IL-4氨基酸序列分別有60%和44%的同源性。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??勾笫驣L-4單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的IL-4會(huì)與單抗結(jié)合形成免疫復(fù)合物,游離的成分被洗去;加入生物素化的抗大鼠IL-4抗體,生物素化抗大鼠IL-4抗體與酶標(biāo)板結(jié)合的標(biāo)準(zhǔn)品或樣本中的大鼠IL-4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-4,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-4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-4濃度。
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