Abstract
Efficient fractionation of peptides is an essential prerequisite for comprehensive analysis of complex protein mixtures by shotgun mass spectrometry. The separation of peptides by isoelectric focusing is particularly attractive due to its orthogonality to reverse-phase HPLC. Here, we present a protocol for in-gel peptide isoelectric focusing using immobilized pH gradient strips. The method shows high resolving power for up to 1 mg of sample and is highly reproducible.
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Acknowledgement
This work was supported by Deutsche Forschungsgemeinschaft (DFG) Grant SFB 958.
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Eravci, M., Sommer, C., Selbach, M. (2014). IPG Strip-Based Peptide Fractionation for Shotgun Proteomics. In: Martins-de-Souza, D. (eds) Shotgun Proteomics. Methods in Molecular Biology, vol 1156. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0685-7_5
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DOI: https://doi.org/10.1007/978-1-4939-0685-7_5
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