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Cells then were lysed in an appropriate amount of CelLytic™ MT Cell Lysis Reagent (Sigma-Aldrich, C3228) containing 1x protease inhibitor cocktail (Sigma-Aldrich, p8340) on ice with agitation for 30 minutes. |
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Protocol tips |
Cells then were lysed in an appropriate amount of CelLytic™ MT Cell Lysis Reagent (Sigma-Aldrich, C3228) containing 1x protease inhibitor cocktail (Sigma-Aldrich, p8340) on ice with agitation for 30 minutes. |
Publication protocol
The appropriate cells were collected by centrifugation at 150 g for 5 minutes after trypsinization and washed once with pre-warmed PBS. Cells then were lysed in an appropriate amount of CelLytic™ MT Cell Lysis Reagent (Sigma-Aldrich, C3228) containing 1x protease inhibitor cocktail (Sigma-Aldrich, p8340) on ice with agitation for 30 minutes. Cell lysates were added to Laemmli sample buffer and boiled for 5 min. Samples were separated on a precast 10% Bis-Tris SDS-PAGE gel.
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The kinetochore-dependent and -independent formation of the CDC20-MAD2 complex and its functions in HeLa cells
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