產(chǎn)品概述
基質(zhì)金屬蛋白酶(MMPs)是鋅依賴性肽鏈內(nèi)切酶,它促進(jìn)細(xì)胞外基質(zhì)蛋白降解,從而控制某些過程的進(jìn)展,如組織構(gòu)建,傷口愈合和瘤轉(zhuǎn)移?;|(zhì)金屬蛋白酶的活性受控于表達(dá)和分泌的調(diào)節(jié),蛋白酶原的活性和基質(zhì)金屬蛋白酶組織抑制劑(TIMPs)。金屬蛋白酶抑制劑通常對金屬蛋白酶具有高度特異性,而對某個(gè)特定的蛋白不是特意的。TIMP-1是可誘導(dǎo)蛋白,TIMP-2是構(gòu)建蛋白,TIMP-3局限在細(xì)胞外基質(zhì),TIMP-4主要局限在心臟組織。TIMP-1是一個(gè)含有184個(gè)氨基酸殘基的糖基化蛋白。TIMP-1可被很多細(xì)胞和組織合成。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薚IMP-1單抗包被于酶標(biāo)板上,同時(shí)加入標(biāo)本、標(biāo)準(zhǔn)品和生物素化的抗人TIMP-1抗體,標(biāo)本、標(biāo)準(zhǔn)品中的TIMP-1會與加入于孔中的生物素化的抗人TIMP-1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TIMP-1,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, TIMP-1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TIMP-1濃度。
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