Organelle Proteomics by Edein P. Romijn, John R. Yates III (auth.), Delphine

By Edein P. Romijn, John R. Yates III (auth.), Delphine Pflieger, Jean Rossier (eds.)

Proteomic analyses have confirmed to be a useful instrument in acquiring high-throughput protein identity from low-abundance, complicated organic samples. In Organelle Proteomics, certain protocols offer step by step directions to effectively learn organelle proteomes by means of appearing the purification of many of the organelles found in eukaryotic cells, in addition to via getting ready convinced sub-fractions of organelles. a sequence of chapters hide the full analytical process of organelle characterization, from its purification beginning with entire cells as much as protein identity utilizing mass spectrometry. dedicated to tools permitting an international estimate of the reliability of the protein checklist assigned to an organelle, Organelle Proteomics permits scientists to achieve an important and critical knowing of organelle study.

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Et al. (2003) Characterization of the human heart mitochondrial proteome. Nat. Biotechnol. 21, 281–286. 15. Washburn, M. , and Yates, J. R. (2001) Large-scale analysis of the yeast proteome by multidimensional protein identification technology. Nat. Biotechnol. 19, 242–247. 16. Wolters, D. , Washburn, M. , and Yates, J. R. (2001) An automated multidimensional protein identification technology for shotgun proteomics. Anal. Chem. 73, 5683–5690. 17. Link, A. J. (2002) Multidimensional peptide separations in proteomics.

22 surgical blades (Feather, Osaka, JP) in dissecting knife handle no. 4 (Fisher, Hampton, NH). 6. 7, and protease inhibitor cocktail in dimethylsulfoxide (DMSO) (250 μL/50 mL buffer) (all from Sigma). 7. , Vineland, NJ). 8. Polytron PT10/35 with PTA 10TS and 12 mm generator (Brinkmann, Westbury, NY). 2. Stacked Golgi Isolation 1. 7 (Sigma), stable at 4°C for up to 1 month. 2. 0 M MgCl2 (Sigma). 3. 0 M sucrose (Sigma): stable at 4°C for up to 1 month. 4. Protease inhibitor cocktail (Sigma). 5. Ultra-Clear 14-mL centrifuge tubes (SW40; Beckman Coulter, Fullerton, CA, USA).

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