Abstract,
Good control, efficiency, and reproducibility of protein extraction from cells and tissues are essential aspects for diverse biological and basic research applications. Effective and specific enzymatic digestion of proteins prior to mass spectrometry (MS) analysis is one of the fundamental techniques most commonly used in any proteomics laboratory. However, neither routine procedures for cell and tissue lysis nor for in-solution and in-gel protein digestion have been significantly altered in common practice for over a decade. Here we have tested and optimized several alternative techniques for preparing lysates of mammalian cells, as well as in-solution and in-gel enzymatic digestion compatible with downstream qualitative and quantitative MS-Based proteomics applications.
摘要,
從細胞和組織提取蛋白質(zhì)的技術(shù)中, 良好的控制,效率和可重復性對于不同生物的分析和基礎研究的應用是一個(gè)非常重要的觀(guān)念。在質(zhì)譜( MS )分析前, 有效和具體的蛋白酶解是一種蛋白質(zhì)組學(xué)實(shí)驗室中zui常用的基本技術(shù)。然而,無(wú)論是細胞和組織溶解的例行程序,或是凝膠蛋白質(zhì)消解, 這十多年已被大大改進(jìn)?;诘鞍踪|(zhì)組學(xué)應用, 我們進(jìn)行了幾個(gè)哺乳動(dòng)物細胞裂解的測試和優(yōu)化, 以及在質(zhì)譜( MS )的基礎下進(jìn)行in-solution and in-gel蛋白酶解的定性和定量分析。
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