Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage
Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage
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DOI：
10.2144/000112401
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发表时间：
2007-04-01
期刊：
影响因子：
2.7
通讯作者：
Ried, Thomas
中科院分区：
文献类型：
--
作者：
Hummon, Amanda B.;Lim, Sharlene R.;Ried, Thomas
系统方法正在生物科学的许多领域得到应用，特别是在癌症研究中。从单个样本中协调、同时提取DNA、RNA和蛋白质对于基因组异常及其对转录组和蛋白质组的影响之间的准确关联至关重要。我们提出了一种提取并完全溶解TIP的方法，该方法从TRIZOL(R)分离RNA和DNA后的存档样本中回收总蛋白的98%。我们还利用聚丙烯酰胺凝胶电泳法(PAGE)和蛋白质印迹分析证明，这些蛋白质在-20摄氏度下在苯酚-乙醇中保持稳定3年，这些蛋白质代表了广泛的相对分子质量范围和一些翻译后修饰，如蛋白质磷酸化。
A systems approach is being applied in many areas of the biological sciences, particularly in cancer research. The coordinated, simultaneous extraction of DNA, RNA, and proteins from a single sample is crucial for accurate correlations between genomic aberrations and their consequences on the transcriptome and proteome. We present an approach to extract and completely solubilize tip to 98% of the total protein recovered from archived samples following TRIZOL (R) isolation of RNA and DNA. We also demonstrate using polyacrylamide gel electrophoresis (PAGE) and Western blot analysis that the proteins, representing both a wide molecular weight range and some posttranslational modifications, such as protein phosphorylation, remain stable in phenol-ethanol for tip to 3 years at -20 degrees C.