Abstract
Sample preparation is a key step in proteomics workflows. Tube-gel (TG) is a fast and repeatable sample preparation method that consists in the instantaneous trapping of the sample in a polyacrylamide gel matrix. It takes advantage of in-gel sample preparations by allowing the use of high concentrations of sodium-dodecyl sulfate but avoids the time-consuming step of electrophoresis. Therefore, TG limits the sample handling and is thus particularly suitable for high-throughput quantitative proteomics when large sample numbers have to be processed, as it is often the case in biomarker research and clinical proteomics projects.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lu X, Zhu H (2005) Tube-gel digestion: a novel proteomic approach for high throughput analysis of membrane proteins. Mol Cell Proteomics 4(12):1948–1958. https://doi.org/10.1074/mcp.M500138-MCP200
An M, Dai J, Wang Q et al (2010) Efficient and clean charge derivatization of peptides for analysis by mass spectrometry. Rapid Commun Mass Spectrom 24(13):1869–1874. https://doi.org/10.1002/rcm.4589
Cao L, Clifton JG, Reutter W et al (2013) Mass spectrometry-based analysis of rat liver and hepatocellular carcinoma Morris hepatoma 7777 plasma membrane proteome. Anal Chem 85(17):8112–8120. https://doi.org/10.1021/ac400774g
Cao R, He Q, Zhou J et al (2008) High-throughput analysis of rat liver plasma membrane proteome by a nonelectrophoretic in-gel tryptic digestion coupled with mass spectrometry identification. J Proteome Res 7(2):535–545. https://doi.org/10.1021/pr070411f
Han CL, Chien CW, Chen WC et al (2008) A multiplexed quantitative strategy for membrane proteomics: opportunities for mining therapeutic targets for autosomal dominant polycystic kidney disease. Mol Cell Proteomics 7(10):1983–1997. https://doi.org/10.1074/mcp.M800068-MCP200
Smolders K, Lombaert N, Valkenborg D et al (2015) An effective plasma membrane proteomics approach for small tissue samples. Sci Rep 5:10917. https://doi.org/10.1038/srep10917
Yu H, Wakim B, Li M et al (2007) Quantifying raft proteins in neonatal mouse brain by ‘tube-gel’ protein digestion label-free shotgun proteomics. Proteome Sci 5:17. https://doi.org/10.1186/1477-5956-5-17
Zhou J, Xiong J, Li J et al (2010) Gel absorption-based sample preparation for the analysis of membrane proteome by mass spectrometry. Anal Biochem 404(2):204–210. https://doi.org/10.1016/j.ab.2010.05.013
Muller L, Fornecker L, Van Dorsselaer A et al (2016) Benchmarking sample preparation/digestion protocols reveals tube-gel being a fast and repeatable method for quantitative proteomics. Proteomics 16(23):2953–2961. https://doi.org/10.1002/pmic.201600288
Acknowledgments
This work was supported by the French Proteomic Infrastructure (ProFI; ANR-10-INBS-08-03).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Muller, L., Fornecker, L., Cianferani, S., Carapito, C. (2019). Tube-Gel: A Fast and Effective Sample Preparation Method for High-Throughput Quantitative Proteomics. In: Brun, V., Couté, Y. (eds) Proteomics for Biomarker Discovery. Methods in Molecular Biology, vol 1959. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9164-8_8
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9164-8_8
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9163-1
Online ISBN: 978-1-4939-9164-8
eBook Packages: Springer Protocols