Pressure-assisted protein extraction: a novel method for recovering proteins from archival tissue for proteomic analysis.
Pressure-assisted protein extraction: a novel method for recovering proteins from archival tissue for proteomic analysis.
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DOI：
10.1021/pr201005t
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发表时间：
2012-04-06
影响因子：
4.4
通讯作者：
Mason, Jeffrey T.
中科院分区：
文献类型：
--
作者：
Fowler, Carol B.;Waybright, Timothy J.;Veenstra, Timothy D.;O'Leary, Timothy J.;Mason, Jeffrey T.
关键词：
甲醛固定，石蜡包埋（FFPE）组织库代表了疾病进展和治疗反应的回顾性研究的宝贵资源。然而，FFPE组织的蛋白质组学分析受到甲醛诱导的蛋白质修饰的阻碍，这降低了蛋白质提取效率并可能导致蛋白质错误鉴定。在这里，我们证明了使用高静水压（40，000 psi）作为一种新的方法，从FFPE小鼠肝脏的完整蛋白质的回收热增强。当FFPE小鼠肝脏使用加热和高压提取时，与仅使用加热提取的匹配组织相比，蛋白质提取效率增加了4倍，完整蛋白质提取增加了3倍，通过质谱鉴定的非冗余蛋白质数量增加了30倍。更重要的是，在FFPE组织中鉴定的非冗余蛋白质的数量与匹配的新鲜冷冻组织几乎相同。
Formaldehyde-fixed, paraffin-embedded (FFPE) tissue repositories represent a valuable resource for the retrospective study of disease progression and response to therapy. However, the proteomic analysis of FFPE tissues has been hampered by formaldehyde-induced protein modifications, which reduce protein extraction efficiency and may lead to protein misidentification. Here, we demonstrate the use of heat augmented with high hydrostatic pressure (40,000 psi) as a novel method for the recovery of intact proteins from FFPE mouse liver. When FFPE mouse liver was extracted using heat and elevated pressure, there was a 4-fold increase in protein extraction efficiency, a 3-fold increase in the extraction of intact proteins, and up to a 30-fold increase in the number of nonredundant proteins identified by mass spectrometry, compared to matched tissue extracted with heat alone. More importantly, the number of nonredundant proteins identified in the FFPE tissue was nearly identical to that of matched fresh-frozen tissue.
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DOI：
10.1007/978-1-59745-324-0_13
发表时间：
2010-01-01
期刊：
IMMUNOCYTOCHEMICAL METHODS AND PROTOCOLS, THIRD EDITION
影响因子：
--
作者：
Bratthauer, Gary L.
通讯作者：
Bratthauer, Gary L.
影响因子：
2.1
作者：
Richardson, Tim;McCanse, Webb;Hemstreet, George P.
通讯作者：
Hemstreet, George P.
影响因子：
3.7
作者：
Fowler CB;Chesnick IE;Moore CD;O'Leary TJ;Mason JT
通讯作者：
Mason JT
影响因子：
3.2
作者：
Guo, Tong;Wang, Weijie;Balgley, Brian M.
通讯作者：
Balgley, Brian M.
影响因子：
7
作者：
Hood, BL;Darfler, MM;Krizman, DB
通讯作者：
Krizman, DB