Abstract
Recently, we reported a new way of performing 2-DE, called P-dimensional electrophoresis (2-PE). In this approach, the second dimension is achieved in a radial gel which can accommodate up to six 7 cm long IPG strips simultaneously, improving reproducibility and throughput power in respect to 2-DE. Nevertheless, 2-PE was up to now limited to the use of only short strips because of technical difficulties. Here, we describe how to load longer strips (e.g., 18–24 cm) on 2-PE and report some representative images for a qualitative assessment.
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References
Campostrini N, Areces LB, Rappsilber J, Pietrogrande MC, Dondi F, Pastorino F, Ponzoni M, Righetti PG (2005) Spot overlapping in two-dimensional maps: a serious problem ignored for much too long. Proteomics 5(9):2385–2395
Lange S, Rosenkrands I, Stein R, Andersen P, Kaufmann SH, Jungblut PR (2013) Analysis of protein species differentiation among mycobacterial low-Mr-secreted proteins by narrow pH range Immobiline gel 2-DE-MALDI-MS. J Proteomics. doi:10.1016/j.jprot.2013.06.036 (Epub ahead of print)
Lee K, Pi K (2009) Effect of separation dimensions on resolution and throughput using very narrow-range IEF for 2-DE after solution phase isoelectric fractionation of a complex proteome. J Sep Sci 32(8):1237–1242. doi:10.1002/jssc.200800672
Lee K, Pi K, Lee HG (2008) Influence of gel dimensions on resolution and sample throughput on two-dimensional gels. Biochemistry 73(4):427–431
Millioni R, Miuzzo M, Antonioli P, Sbrignadello S, Iori E, Dosselli R, Puricelli L, Kolbe M, Tessari P, Righetti PG (2010a) SDS-PAGE and two-dimensional maps in a radial gel format. Electrophoresis 31(3):465–470. doi:10.1002/elps.200900526
Millioni R, Miuzzo M, Puricelli L, Iori E, Sbrignadello S, Dosselli R, Cecconi D, Tessari P, Righetti PG (2010b) Improved instrumentation for large-size two-dimensional protein maps. Electrophoresis 31(23–24):3863–3866. doi:10.1002/elps.201000460
Millioni R, Polati R, Menini M, Puricelli L, Miuzzo M, Tessari P, Novelli E, Righetti PG, Cecconi D (2012) Polar electrophoresis: shape of two-dimensional maps is as important as size. PLoS One 7(1):e30911. doi:10.1371/journal.pone.0030911
Minden JS, Dowd SR, Meyer HE, Stühler K (2009) Difference gel electrophoresis. Electrophoresis 30(Suppl 1):S156–S161. doi:10.1002/elps.200900098
Oguri T, Takahata I, Katsuta K, Nomura E, Hidaka M, Inagaki N (2002) Proteome analysis of rat hippocampal neurons by multiple large gel two-dimensional electrophoresis. Proteomics 2:666–672
Poland J, Cahill MA, Sinha P (2003) Isoelectric focusing in long immobilized pH gradient gels to improve protein separation in proteomic analysis. Electrophoresis 24(7–8):1271–1275
Polati R, Menini M, Robotti E, Millioni R, Marengo E, Novelli E, Balzan S, Cecconi D (2012) Proteomic changes involved in tenderization of bovine Longissimus dorsi muscle during prolonged ageing. Food Chem 135(3):2052–2069. doi:10.1016/j.foodchem.2012.06.093
Righetti PG (1983) Isoelectric focusing: theory, methodology and applications. Elsevier, Amsterdam
Yuan Q, An J, Liu DG, Zhao FK (2003) Multi-strips on one gel method to improve the reproducibility, resolution power and high-throughput of two-dimensional electrophoresis. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao 35(7):611–618
Acknowledgments
The authors greatly thank prof. Pier Giorgio Righetti, Dr. Paolo Antonioli, and Antonio Guadagnino for their support in determining the experimental setting of P-dimensional electrophoresis. The authors greatly thank also Elettrofor SAS, Eletech SRL, Zanchi Investimenti e Partecipazioni SRL for supporting the study and the instrumental development.
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The authors have declared no conflict of interest.
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P. Rita and C. Daniela have equally contributed to this work.
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Polati, R., Cecconi, D., Dosselli, R. et al. Improvements to polar 2-D electrophoresis for proteomic applications. Amino Acids 46, 1143–1146 (2014). https://doi.org/10.1007/s00726-014-1672-8
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DOI: https://doi.org/10.1007/s00726-014-1672-8