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Quantifying raft proteins in neonatal mouse brain by 'tube-gel' protein digestion label-free shotgun proteomics |
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| Authors: | Hongwei Yu Bassam Wakim Man Li Brian Halligan G Stephen Tint Shailendra B Patel |
| Affiliation: | (1) Division of Endocrinology, Metabolism and Nutrition, Medical College of Wisconsin, Milwaukee, WI 53226, USA;(2) Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA;(3) National Center for Proteomics Research, Biotechnology and Bioinformatics Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA;(4) Research Service, Department of Veterans Affairs, New Jersey Health Care System, East Orange, NJ 07018, USA;(5) Department of Medicine, UMDNJ-New Jersey Medical School, Newark, NJ 07103-2714, USA;(6) Department of Veterans Affairs, Clement J. Zablocki Medical Center, Milwaukee, WI 53295, USA;(7) Qilu Hospital, Shandong University, 44 West Wenhua Road, Jinan, 250012, P. R., China |
| Abstract: | BackgroundThe low concentration and highly hydrophobic nature of proteins in lipid raft samples present significant challenges for the sensitive and accurate proteomic analyses of lipid raft proteins. Elimination of highly enriched lipids and interfering substances from raft samples is generally required before mass spectrometric analyses can be performed, but these procedures often lead to excessive protein loss and increased sample variability. For accurate analyses of the raft proteome, simplified protocols are needed to avoid excessive sample handling and purification steps. |
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