Protein extraction from formalin-fixed, paraffin-embedded tissue sections: Quality evaluation by mass spectrometry
Protein extraction from formalin-fixed, paraffin-embedded tissue sections: Quality evaluation by mass spectrometry
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DOI：
10.1369/jhc.5b6851.2006
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发表时间：
2006-06-01
影响因子：
3.2
通讯作者：
Taylor, Clive R.
中科院分区：
文献类型：
--
作者：
Shi, Shan-Rong;Liu, Cheng;Taylor, Clive R.
基于广泛应用于免疫组织化学中档案福尔马林固定石蜡包埋（FFPE）组织切片的热诱导抗原修复（AR）技术，建立了令人满意的蛋白质提取方案。基于 AR，使用 FFPE 小鼠组织进行了初步系列实验，以确定热诱导蛋白提取的最佳方案。然后在人类肾癌的档案 FFPE 组织上执行提取蛋白质的最佳方案。 FFPE 切片在含有 2% SDS 的 Tris-HCl 修复液中煮沸，然后孵育。采用放射免疫沉淀法对同例肾癌新鲜组织进行常规方法提取蛋白，比较FFPE组织切片与新鲜组织提取蛋白的效率。作为对照，同一 FFPE 样品的其他部分通过相同的程序进行处理，无需进行热处理。使用凝胶电泳和质谱法对从 FFPE 组织中提取的蛋白质的质量进行了评估，结果显示从 FFPE 组织切片中提取的大多数已鉴定蛋白质与从新鲜组织中提取的蛋白质重叠。
A satisfactory protocol of protein extraction has been established based on the heat-induced antigen retrieval (AR) technique widely applied in immunohistochemistry for archival formalin-fixed, paraffin-embedded (FFPE) tissue sections. Based on AR, an initial serial experiment to identify an optimal protocol of heat-induced protein extraction was carried out using FFPE mouse tissues. The optimal protocol for extraction of proteins was then performed on an archival FFPE tissue of human renal carcinoma. FFPE sections were boiled in a retrieval solution of Tris-HCI containing 2% SDS, followed by incubation. Fresh tissue taken from the same case of renal carcinoma was processed for extraction of proteins by a conventional method using radioimmunoprecipitation assay solution, to compare the efficiency of protein extraction from FFPE tissue sections with extraction from fresh tissue. As a control, further sections of the same FFPE sample were processed by the same procedure without heating treatment. Evaluation of the quality of protein extracted from FFPE tissue was done using gel electrophoresis and mass spectrometry, showing most identified proteins extracted from FFPE tissue sections were overlapped with those extracted from fresh tissue.