Dramatic improvement of proteomic analysis of zebrafish liver tumor by effective protein extraction with sodium deoxycholate and heat denaturation.
Dramatic improvement of proteomic analysis of zebrafish liver tumor by effective protein extraction with sodium deoxycholate and heat denaturation.
复制标题
DOI：
10.1155/2015/763969
复制
发表时间：
2015
影响因子：
1.8
通讯作者：
Lin Q
中科院分区：
文献类型：
--
作者：
Wang J;Lee YM;Li C;Li P;Li Z;Lim TK;Gong Z;Lin Q
大多数组织样本的蛋白质组学研究涉及使用基于凝胶的方法进行分析和消化。费力的基于凝胶的方法正在慢慢被先进的溶液中消化方法所取代。然而，仍然存在一些困难，如难以溶解的蛋白质，在复杂组织样品中的蛋白质组学分析差，以及样品杂质的存在。然而，对配制高效蛋白质提取缓冲液存在很大需求，所述缓冲液具有从组织样品的高蛋白质提取效率、与溶液内消化的高相容性、减少样品处理步骤的数量以减少样品损失、低时间消耗、低成本和易于使用。在这里，我们评估了各种现有的蛋白质提取缓冲液与斑马鱼肝肿瘤样品，发现脱氧胆酸钠（DOC-）为基础的提取缓冲液与热变性是最有效的方法，从复杂的组织，如斑马鱼肝肿瘤蛋白质的高效提取。共鉴定了4,790个蛋白质，使用鸟枪蛋白质组学方法与2D LC，据我们所知，这是最全面的研究斑马鱼肝肿瘤蛋白质组。
Majority of the proteomic studies on tissue samples involve the use of gel-based approach for profiling and digestion. The laborious gel-based approach is slowly being replaced by the advancing in-solution digestion approach. However, there are still several difficulties such as difficult-to-solubilize proteins, poor proteomic analysis in complex tissue samples, and the presence of sample impurities. Henceforth, there is a great demand to formulate a highly efficient protein extraction buffer with high protein extraction efficiency from tissue samples, high compatibility with in-solution digestion, reduced number of sample handling steps to reduce sample loss, low time consumption, low cost, and ease of usage. Here, we evaluated various existing protein extraction buffers with zebrafish liver tumor samples and found that sodium deoxycholate- (DOC-) based extraction buffer with heat denaturation was the most effective approach for highly efficient extraction of proteins from complex tissues such as the zebrafish liver tumor. A total of 4,790 proteins have been identified using shotgun proteomics approach with 2D LC, which to our knowledge is the most comprehensive study for zebrafish liver tumor proteome.
登录
查看更多内容
影响因子：
2.9
作者：
GORDON, JA;JENCKS, WP
通讯作者：
JENCKS, WP
影响因子：
46.9
作者：
Washburn, MP;Wolters, D;Yates, JR
通讯作者：
Yates, JR
影响因子：
7.4
作者：
Bhatia, Vivek N.;Perlman, David H.;Costello, Catherine E.;McComb, Mark E.
通讯作者：
McComb, Mark E.
影响因子：
4.5
作者：
Lachenmayer, Anja;Alsinet, Clara;Chang, Charissa Y.;Llovet, Josep M.
通讯作者：
Llovet, Josep M.
影响因子：
11.5
作者：
Sahin, F;Kannangai, R;Torbenson, M
通讯作者：
Torbenson, M