Translational Cancer Research Group, Proteo-Red-ISCIII, Instituto de Investigación Biomédica A Coruña, Coruña, Spain.
Proteomics. 2011 Jun;11(12):2555-9. doi: 10.1002/pmic.201000809. Epub 2011 May 18.
Most of the archived pathological specimens in hospitals are kept as formalin-fixed paraffin-embedded tissues (FFPE) for long-term preservation. Up to now, these samples are only used for immunohistochemistry in a clinical routine as it is difficult to recover intact protein from these FFPE tissues. Here, we report a novel, short time-consuming and cost-effective method to extract full-length, non-degraded proteins from FFPE tissues. This procedure is combined with an effective and non-toxic deparaffinisation process and an extraction method based on antigen-retrieval, high concentration of SDS and high temperature. We have obtained enough intact protein to be detected by Western blotting analysis. This technique will allow utilising these stored FFPE tissues in several applications for protein analysis helping to advance the translational studies in cancer and other diseases.
大多数医院存档的病理标本都以福尔马林固定石蜡包埋组织(FFPE)的形式长期保存。到目前为止,这些样本仅在临床常规中用于免疫组织化学,因为从这些 FFPE 组织中回收完整的蛋白质非常困难。在这里,我们报告了一种从 FFPE 组织中提取全长、无降解蛋白质的新型、耗时短且经济高效的方法。该方法结合了一种有效且无毒的脱蜡过程以及一种基于抗原回收、高浓度 SDS 和高温的提取方法。我们已经获得了足够的完整蛋白质,可以通过 Western blot 分析进行检测。该技术将允许在蛋白质分析的多个应用中利用这些储存的 FFPE 组织,从而有助于推进癌症和其他疾病的转化研究。