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Biochemistry

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JoVE Journal Biochemistry
Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
 

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Article DOI: 10.3791/59079-v • 11:54 min • March 23rd, 2020
March 23rd, 2020 •
1, 1, 1
1Proteomics Center, Erasmus University Medical Center

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We present a method for the purification, detection, and identification of diGly peptides that originate from ubiquitinated proteins from complex biological samples. The presented method is reproducible, robust, and outperforms published methods with respect to the level of depth of the ubiquitinome analysis.

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Protein Ubiquitination Sites Peptide Enrichment Mass Spectrometry Post Translational Modification DiGly Peptides Crude Peptide Fractionation Orbitrap Ubiquitinome Visualization Cultured Cells Mouse Brain Tissue Lyse Tris HCL Sodium Deoxycholate
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