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明膠酶譜分析試劑盒組成

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1. 簡介: 
基質(zhì)金屬蛋白酶(matrix metalloproteinase, MMP)以無活性的酶原形式分泌,活化后能夠降解細(xì)胞外基質(zhì)的膠原蛋白,又稱膠原酶。通過影響細(xì)胞外基質(zhì),膠原酶參與胚胎發(fā)育、形態(tài)發(fā)生、組織重塑等過程,并參與炎癥、腫瘤、心血管、神經(jīng)等許多疾病過程。血漿與組織中的MMP-2、MMP-9參與各種生理與病理過程,其在診斷和治療中的意義日益受到重視。

2. 檢測原理: 
本試劑盒采用酶譜法(Zymography)檢測MMP-2、MMP-9 活性。Zymography 是一種廣為使用的、基于SDS-PAGE 電泳和反相凝膠染色的蛋白酶檢測方法,其靈敏度可達(dá)1 nM,高于ELISA方法,其基本原理和程序是:制備加有膠原酶底物的SDS-PAGE 凝膠,含蛋白酶的樣品在此凝膠中進(jìn)行電泳,電泳結(jié)束后取出凝膠與酶反應(yīng)Buffer 孵育,凝膠染色與脫色。凝膠中由于含底物蛋白而被深染,形成深色背景,但在有蛋白酶條帶的位置,蛋白酶將降解凝膠中的底物蛋白,因而不被染色而形成透亮區(qū)域,從而能同時(shí)指示MMP-2、MMP-9 的大小位置(酶譜)及活性。本試劑盒提供MMP-2、MMP-9 特異蛋白底物及酶學(xué)反應(yīng)緩沖液,可檢測少至0.1~0.5 μl 血液中的MMP-2、MMP-9 酶譜及活性,檢測靈敏度~1nM。試劑盒可進(jìn)行50次標(biāo)準(zhǔn)小膠檢測,如果小膠加樣孔為10~15個(gè),則總計(jì)可檢測500~750個(gè)樣品。

 

參考文獻(xiàn):
1. Nagase H and Woessner JF. Matrix Metalloproteinases. J Biol Chem, 1999, 274, 21491-21494
2. Heussen C, and Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide
gels containing sodium dodecyl sulfate and copolymerized substrates. Anal. Biochem,1980, 
102,196–202

 
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