產(chǎn)品概述
基質(zhì)金屬蛋白酶(MMPs)是鋅依賴性降解細(xì)胞外基質(zhì)蛋白的內(nèi)肽酶家族中的一員。它以酶原(pro-MMPs)形式被分泌,可以激活多種蛋白酶或是在有機(jī)汞存在的條件下發(fā)揮作用。MMP的活性調(diào)控在組織重建、炎癥、腫瘤生成和轉(zhuǎn)移中起重要作用。單核細(xì)胞、巨噬細(xì)胞、嗜中性粒細(xì)胞、角化細(xì)胞、成纖維細(xì)胞、成骨細(xì)胞、軟骨細(xì)胞、骨骼肌星狀細(xì)胞、血管內(nèi)皮細(xì)胞和各種腫瘤細(xì)胞都可以分泌酶原 MMP -9。TGF-β 1,IL-1β,TGF-α,PDGF-AB,TNF-α和IL-1α可以使酶原MMP-9表達(dá)上調(diào)。 MMP-9 與炎癥反應(yīng)、組織構(gòu)建、傷口愈合、生長(zhǎng)因子和細(xì)胞因子過(guò)程有關(guān)。它的表達(dá)與伴隨胰腺癌生成的結(jié)締組織、與人乳腺癌細(xì)胞轉(zhuǎn)移到淋巴結(jié)和骨巨噬細(xì)胞瘤入侵局部導(dǎo)管相關(guān)。 MMP-9 在齦溝液、齒齦炎、牙周炎患者的唾液可能表達(dá)比較高。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薓MP-9單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的MMP-9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MMP-9抗體,生物素化抗人MMP-9抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的MMP-9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MMP-9,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, MMP-9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MMP-9濃度。
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