產(chǎn)品概述
單核細(xì)胞趨化蛋白1(MCP-1/CCL2),也稱為單核細(xì)胞趨化與激活因子(MCAF),是趨化因子家族CC亞族一個(gè)成員。MCP-1是一種很有潛能的單核細(xì)胞及巨噬細(xì)胞的誘生劑及激活因子。人MCP-1 cDNA編碼99個(gè)氨基酸組成的前體蛋白,當(dāng)剪切掉23個(gè)氨基酸疏水性信號(hào)肽后,就形成了由76個(gè)氨基酸組成的成熟蛋白。與小鼠趨化因子相比,人MCP-1與小鼠MCP-5(CCL12)、MCP-1/JE(CCL2)最相關(guān),小鼠MCP-5與人MCP-1有65%序列同源性。MCP-1可以被許多炎癥介導(dǎo)物誘導(dǎo),如LPS、 IL-1β、 TNF-α、IFN-γ、IL-6及IL-4等。表達(dá)MCP-1的細(xì)胞有:?jiǎn)魏思?xì)胞、肥大細(xì)胞、T細(xì)胞、成骨細(xì)胞、成纖維細(xì)胞、內(nèi)皮細(xì)胞、表皮細(xì)胞、小膠質(zhì)細(xì)胞、星形膠質(zhì)細(xì)胞及平滑肌細(xì)胞等。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薓CP-1單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的MCP-1會(huì)與單抗結(jié)合形成免疫復(fù)合物,游離的成分被洗去;加入生物素化的抗人MCP-1抗體,生物素化抗人MCP-1抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的MCP-1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MCP-1,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, MCP-1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MCP-1濃度。
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