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. 2013 Sep 19;8(9):e75309. doi: 10.1371/journal.pone.0075309

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© 2013 la Cour et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A. Recombinant proteins bound to artificial liposomes and collected from a sucrose gradient were analyzed for recruitment efficiency in the presence of ALG-2 and a peptide representing the ABD (pABD). Antibodies directed against Sec31A, HA tag (HA-Sec13), Flag-tag (flag-Sec23A), Sar1 and ALG-2 were used to analyze protein recruitment. The immunoblot is representative of more than three independent experiments. B. Quantification of band intensities relative to Ca²⁺ control (ctr) in representative immunoblot. C. Anti-Flag antibody coated beads were used as bait for flag-Sec23. Protein binding was performed in the presence of Ca²⁺ and proteins in gels were visualized using SyproRed staining.