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[交流]
zymography（酶谱法）？
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我前两天看到两篇关于脂肪酶表达的文献，在用SDS-PAGE法检测所表达的胞外脂肪酶分子量大小时，文中还同时用zymography法检测了其大小，所用的染料为Fast Red TR Salt，具体操作步骤如下： SDS^PAGE and zymography： SDS^PAGE of proteins was performed in a 12% polyacrylamide gel by the method of Laemmli . The transformants of E. coli induced with IPTG were collected by centrifugation, washed with 50 mM acetate bu¡er (pH 5.5), and then suspended in the sample bu¡er. The lysate solution was subjected to SDS^PAGE. To detect esterases including lipases, the gel was soaked at room temperature in a solution containing 1-naphthylcaprylate instead of 1- naphthylacetate (Sigma) and Fast Red TR Salt (Aldrich), and the activity was visualized as a red band . 我想问一下酶谱法的原理（请详细点喔！）及具体到这个实验的原理！虫们帮帮忙啊！ |
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4楼2008-04-09 15:19:14
2楼2008-04-07 22:51:49
glucidum
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3楼2008-04-08 08:47:24