產(chǎn)品概述
γ干擾素(IFN-γ)也被稱為二型干擾素,最初被認(rèn)為是由促有絲分裂原激活的T淋巴細(xì)胞產(chǎn)生具有抗病毒活性物質(zhì)。INF-γ經(jīng)證實(shí)是在主體防御中起重要作用,包括免疫反應(yīng)、反增生和引發(fā)炎癥反應(yīng)。干擾素γ主要通過T淋巴細(xì)胞、自然殺傷細(xì)胞和CD8+毒素細(xì)胞產(chǎn)生。另外一類型細(xì)胞包括嗜中性粒細(xì)胞、柱狀細(xì)胞、角化細(xì)胞、感覺神經(jīng)細(xì)胞、巨噬細(xì)胞和B細(xì)胞,這些細(xì)胞都可以有IFN-γ表達(dá)。大鼠IFN-γcDNA編碼156氨基酸殘基的前體蛋白和具有19個(gè)氨基酸殘基的信號(hào)縮氨酸裂解產(chǎn)生的成熟蛋白,它包含兩個(gè)潛在N端位點(diǎn)。天然的IFN-γ序列的三個(gè)N端(Cys-Tyr-Cys)是分離的。IFN-γ的C末端也有很高的蛋白水解活性。重組和自然產(chǎn)生IFN-γ以非共價(jià)連接的同型二聚體存在于溶液中。大鼠IFN-γ與小鼠IFN-γ的同源性為87%,與人IFN-γ的同源性為39%。大鼠IFN-γ對(duì)小鼠細(xì)胞有活性,對(duì)人類細(xì)胞沒活性。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法。抗大鼠IFN-γ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IFN-γ抗體,標(biāo)本、標(biāo)準(zhǔn)品中的IFN-γ會(huì)與加入于孔中的生物素化的抗大鼠IFN-γ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IFN-γ,辣根過氧化物酶會(huì)使無色的顯色劑變藍(lán),加終止液變黃。在450nm處測OD值,IFN-γ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IFN-γ濃度。
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