A comprehensive evaluation of imidazole‐zinc reverse stain for current proteomic researches |
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| Authors: | Ching‐Yu Lin Vinchi Wang Hao‐Ai Shui Rong‐Huay Juang Ai‐Ling Hour Pei‐Sing Chen Hui‐Ming Huang Szu‐Yu Wu Jen‐Chieh Lee Tzung‐Lin Tsai Han‐Min Chen Professor |
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| Institution: | 1. Graduate Institute of Applied Science and Engineering, Catholic Fu‐Jen University, Taipei, Taiwan, Republic of China;2. Department of Life‐Science, Catholic Fu‐Jen University, Taipei, Taiwan, Republic of China;3. Department of Neurology, Cardinal Tein Hospital, Taipei, Taiwan, Republic of China;4. School of Medicine, Catholic Fu‐Jen University, Taipei, Taiwan, Republic of China;5. Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China;6. Department of Biochemical Science and Technology, and Institute of Microbiology and Biochemistry, National Taiwan University, Taipei, Taiwan, Republic of China |
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| Abstract: | In this paper, we comprehensively evaluated the capability of imidazole‐zinc reverse stain (ZN) in comparative proteomics. Three commonly used protein gel staining methods, including silver (SN), SYPRO Ruby (SR), and CB stain were investigated alongside for comparison purpose. A transparency scanning procedure, which may deliver more even and contrasting gel images, was found best for documenting ZN stained gels. Our results showed that ZN was more sensitive than SN, SR, and CB. It may reveal as few as 1.8 ng of proteins in a gel. Moreover, ZN was found to provide a linear dynamic range of staining for revealing proteins up to 140 ng, and show an insignificant staining preference. To analyze a ZN stained 2‐D gel image that generally comprises an apparent but even background, the Melanie 4 software was found more suitable than others. Furthermore, ZN demonstrated an equivalent or better MS compatibility than the other three staining methods. Intense and comprehensive MS profiles were frequently observed for ZN stained gel spots. Approximate two‐third of ZN stained gel spots were successfully identified for protein identities. Taken together, our results suggest that the prompt, cost effective and versatile ZN is well suited for current proteomic researches. |
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| Keywords: | 2‐DE Evaluation Imidazole‐zinc reverse stain Proteomics |
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