Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.
Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.
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DOI：
10.1002/prca.201200064
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发表时间：
2013-04
影响因子：
2
通讯作者：
Van Eyk, Jennifer E.
中科院分区：
文献类型：
--
作者：
Fu, Zongming;Yan, Kun;Rosenberg, Avraham;Jin, Zhicheng;Crain, Barbara;Athas, Grace;Vander Heide, Richard S.;Howard, Timothy;Everett, Allen D.;Herrington, David;Van Eyk, Jennifer E.
评价加热和高压的组合，以提高蛋白质提取和甲醛固定（FF）和FF石蜡包埋（FFPE）主动脉的质量，用于蛋白质组学。在室温下从新鲜冷冻的主动脉中提取蛋白质。在室温、单独加热或加热和高压组合下提取FF和FFPE动脉瘤（3个月和15年）。比较蛋白质产率，并用质谱法分析来自提取物的消化肽。与单独加热（1.5倍）或RT（8.3倍）的匹配组织相比，组合加热和升高的压力增加了人FF或FFPE主动脉的蛋白质产量，导致鉴定出更多的蛋白质，并具有更多的序列覆盖率。储存时间的长短确实对FF组织蛋白质的质量产生不利影响。对于长期储存，使用FFPE比单独使用FF更好地保存主动脉。证明骨膜蛋白和MGF-E8适用于来自FFPE主动脉的MRM测定。热高压结合是从FFPE主动脉中提取蛋白质用于下游蛋白质组学研究的有效方法。这种方法打开了使用存档的和通常罕见的FFPE组织和可能的其他组织可用于蛋白质组学的生物标志物发现和定量的可能性。
Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formaldehyde-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics. Proteins were extracted from fresh frozen aorta at RT. FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with mass spectrometry. Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5 fold) or at RT (8.3 fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the quality of proteins from FF tissue. For long term storage, aorta was preserved better with FFPE than FF alone. Periostin and MGF-E8 were demonstrated suitable for MRM assays from FFPE aorta. Combination of heat and high pressure is an effective method to extract proteins from FFPE aorta for downstream proteomics. This method opens the possibility for use of archival and often rare FFPE aortas and possibly other tissues available to proteomics for biomarker discovery and quantification.
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DOI：
10.1161/atvbaha.111.224352
发表时间：
2011-07
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Hixson JE;Shimmin LC;Montasser ME;Kim DK;Zhong Y;Ibarguen H;Follis J;Malcom G;Strong J;Howard T;Langefeld C;Liu Y;Rotter JI;Johnson C;Herrington D
通讯作者：
Herrington D
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：
7.3
作者：
Nirmalan, Niroshini J.;Harnden, Patricia;Banks, Rosamonde E.
通讯作者：
Banks, Rosamonde E.
DOI：
10.1074/mcp.m110.001693
发表时间：
2010-09
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Didangelos A;Yin X;Mandal K;Baumert M;Jahangiri M;Mayr M
通讯作者：
Mayr M
影响因子：
4.4
作者：
Ostasiewicz, Pawel;Zielinska, Dorota F.;Wisniewski, Jacek R.
通讯作者：
Wisniewski, Jacek R.