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Fig. 2. Immunoblot analysis of tobacco BY-2 cell extracts. (A) Extracts from cells expressing the Spc98p-GFP fusion protein. Lane 1, anti-{alpha}-tubulin (1/5000); lane 2, anti-ß-tubulin (1/5000); lane 3, anti-{gamma}-tubulin (1/1000), which does not crossreact with {alpha}- or with ß-tubulin; lane 4, anti-GFP (1/2000) reveals one band at the expected size of the fusion protein; lane 5, anti-Spc98pB (1/1000) reveals the endogenous Spc98p (95 kDa, double arrow) as well as the fusion protein (125 kDa, arrow). (B) Lanes 6 to 8, affinity-purified antibodies directed against three Spc98 peptides (pA, pB and pC) were tested on control BY-2 extracts; best results were obtained using the pB peptide (DLDSIAKDYTSSLDA), a plant/animal consensus sequence. Antibodies directed against the C-terminal peptide pC did not crossreact with BY-2 extracts.





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