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Two-step immunoprecipitation of kinetochores. (A) Workflow for two-step immunoprecipitation using a second immunoprecipitation of Spc24–3V5 following purification of Dsn1-HIS-FLAG. (B) Immunoblot of two-step immunoprecipitation shows full depletion of Spc24–3V5 but not Dsn1-HIS-FLAG. (C) Silver staining of SDS-PAGE of one-step and two-step purified kinetochores.
