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. 2002 Jun;22(12):4167–4180. doi: 10.1128/MCB.22.12.4167-4180.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 1. — Purification of Sir3 and Sir2/Sir4 from yeast extracts. Silver stained SDS-polyacrylamide gels showing the purification of TAP-tagged Sir3 (A) and Sir4 (B) from strains DMY1737 and DMY1704, respectively. The left panel in A shows a control purification from the parental strain lacking the TAP tag (SF10). Proteins were identified by mass spectrometry. The band marked with an asterisk (∗) in B was identified as Ssb1, a yeast Hsp70 protein, which is a common contaminant of TAP purifications. Lanes 1 to 4 show successive elution fractions from calmodulin-Sepharose, the second column used in the purifications. (C) Western blots showing the migration of Sir3 in a Superose 6 gel filtration column in crude yeast extracts (Sir3-TAP, top panel) and after purification on IgG-Sepharose and calmodulin-Sepharose (Sir3-CBP, top panel, same material as in A, lane 3). Molecular size markers used for calibration of the sizing column were thyroglobulin (670 kDa) and aldolase (158 kDa).