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Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80

Article DOI: 10.3791/52482-v • 12:28 min • May 3rd, 2015
May 3rd, 2015 •
1, 1, 1
1Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institute of Health

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This manuscript describes the detection of sumoylation and ubiquitination of kinetochore proteins, Ndc10 and Ndc80, in the budding yeast Saccharomyces cerevisiae.

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Protein Purification Technique Detection Sumoylation Ubiquitination Budding Yeast Kinetochore Proteins Ndc10 Ndc80 Post-translational Modifications PTMs Phosphorylation Methylation Acetylation Mass Spectrometry Western Blot Analysis Biochemical Approaches Genome Stability Saccharomyces Cerevisiae Polyhistidine-Flag-tagged Smt3 Myc-tagged Ndc10 Myc-tagged Ndc80 Affinity Purification Nickel Beads Anti-Myc Antibody Anti-Ub Antibody Epitope Tagged-protein
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