Evaluation of Optimized Tube-Gel Methods of Sample Preparation for Large-Scale Plant Proteomics
AbstractThe so-called tube-gel method is a sample preparation protocol allowing for management of SDS for protein solubilization through in-gel protein trapping. Because of its simplicity, we assumed that once miniaturized, this method could become a standard for large scale experiments. We evaluated the performances of two variants of the miniaturized version of the tube-gel method based on different solubilization buffers (Tris-SDS or urea-SDS). To this end, we compared them to two other digestion methods: (i) liquid digestion after protein solubilization in the absence of SDS (liquid method) and (ii) filter-aided sample preparation (FASP). As large-scale experiments may require long term gel storage, we also examined to which extent gel aging affected the results of the proteomics analysis. We showed that both tube-gel and FASP methods extracted membrane proteins better than the liquid method, while the latter allowed the identification and quantification of a greater number of proteins. All methods were equivalent regarding quantitative stability. However, important differences were observed regarding post-translational modifications. In particular, methionine oxidation was higher with the tube-gel method than with the other methods. Based on these results, and considering time, simplicity, and cost aspects, we conclude that the miniaturized tube-gel method is suitable for sample preparation in the context of large-scale experiments. View Full-Text
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Balliau, T.; Blein-Nicolas, M.; Zivy, M. Evaluation of Optimized Tube-Gel Methods of Sample Preparation for Large-Scale Plant Proteomics. Proteomes 2018, 6, 6.
Balliau T, Blein-Nicolas M, Zivy M. Evaluation of Optimized Tube-Gel Methods of Sample Preparation for Large-Scale Plant Proteomics. Proteomes. 2018; 6(1):6.Chicago/Turabian Style
Balliau, Thierry; Blein-Nicolas, Mélisande; Zivy, Michel. 2018. "Evaluation of Optimized Tube-Gel Methods of Sample Preparation for Large-Scale Plant Proteomics." Proteomes 6, no. 1: 6.
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