Lauber W M, Carroll J A, Dufield D R, Kiesel J R, Radabaugh M R, Malone J P
Pharmacia Corp, St Louis, MO 63198, USA.
Electrophoresis. 2001 Mar;22(5):906-18. doi: 10.1002/1522-2683()22:5<906::AID-ELPS906>3.0.CO;2-9.
As proteomic technology evolves, protein staining sensitivity is constantly being improved, enabling researchers to better visualize the proteome of their system. The current challenge is to balance the limits of detection of protein visualization with those of the mass spectrometric methods. In this report, mass spectra generated from human serum or rat liver proteins stained with either colloidal Coomassie blue, Daiichi silver, SYPRO Orange, SYPRO Red, SYPRO Ruby, or SYPRO Tangerine are compared. It has been concluded that the newest generation of fluorescent protein stains, compared with traditional staining methods, are more compatible to matrix-assisted laser desorption/ionization (MALDI) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. The number of database matches obtained using each mass spectrometry method and the percent sequence coverage obtained from trypsin digested proteins stained using these six methods is provided.
随着蛋白质组学技术的发展,蛋白质染色灵敏度不断提高,使研究人员能够更好地观察其系统的蛋白质组。当前的挑战是要在蛋白质可视化检测极限与质谱方法检测极限之间取得平衡。在本报告中,对用考马斯亮蓝胶体、第一银染、SYPRO Orange、SYPRO Red、SYPRO Ruby或SYPRO Tangerine染色的人血清或大鼠肝脏蛋白质所产生的质谱进行了比较。得出的结论是,与传统染色方法相比,最新一代的荧光蛋白质染色剂与基质辅助激光解吸/电离(MALDI)和液相色谱-串联质谱(LC-MS/MS)方法更兼容。提供了使用每种质谱方法获得的数据库匹配数以及用这六种方法染色的经胰蛋白酶消化的蛋白质的序列覆盖率百分比。