By Herve, Ed. Thiellement
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Additional info for Plant Proteomics: Methods and Protocols (Methods in Molecular Biology Vol 355)
Lucas, W. J. (1999). Plasmodesmata and the cell-to-cell transport of proteins and nucleoprotein complexes. J. Exp. Bot. 50, 979–987. 13. , and Fisahn, J. (1999). Analysis of phloem protein patterns from different organs of Cucurbita maxima Duch. by matrix-assisted laser desorption/ionisation time of flight mass spectroscopy combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Planta 207, 612–619. 14. , and Lucas, W. J. (2002). Analysis of the complexity of protein kinases within the phloem sieve tube system.
5. Add to 250 µL of the supernatant 20 µL of carrier ampholytes (according to the IPG strip gradient used in the IEF step) and 4 µL 4-vinylpyridine (see Note 11). After a contact of 15 min and vortexing, 254 µL of the glycerol solution is added (see Note 12). The protein extract can be stored at –80°C until use or analyzed directly in 2-DE. 2. , water-soluble albumins and salt-soluble globulins) has been optimized for the IEF. Thus, after being extracted with the sodium-phosphate buffer as described elsewhere (18), the protein extract is desalted and the proteins are then precipitated and solubilized in a buffer compatible with the IEF.
Wong, J. , Buchanan, B. , and Hurkman, W. J. (2005) Developmental changes in the metabolic protein profiles of wheat endosperm. Proteomics 5, 1594–1611 13. Wong, J. , et al. (2004) Thioredoxin targets of developing wheat seeds identified by complementary proteomic approaches. Phytochemistry 65, 1629–1640. 14. , and Svensson, B. (2002) Initial proteome analysis of mature barley seeds and malt. Proteomics 2, 733–739. 15. , and Svensson, B. (2004) Proteome analysis of barley seeds: identification of major proteins from twodimensional gels (pI 4-7).