A parallel affinity purification method for selective isolation of polyubiquitinated proteins
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Updated: Jul 3, 2026

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations
Published on: November 7, 2019
Kazuhisa Ota1, Keiji Kito, Shun-ichiro Iemura
1Department of Computational Biology, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan.
Abstract:
We developed a parallel affinity purification (PAP) procedure, in which ubiquitinated proteins are purified from the cells that coexpress two affinity-tagged ubiquitins by sequential use of affinity chromatography specific to each tag. In contrast with previous procedures using a single affinity-tagged ubiquitin, the PAP eliminates highly abundant ubiquitin monomers and monoubiquitinated proteins to selectively enrich proteins bearing both affinity-tags, or poly- and multiubiquitinated proteins. Accordingly, it would serve as a powerful method to facilitate mass-spectrometric identification of ubiquitinated proteins.

