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. 2013 Dec 15;443(2):153-5.
doi: 10.1016/j.ab.2013.09.011. Epub 2013 Sep 16.

Simultaneous quantification of total and conjugated ubiquitin levels in a single immunoblot

Choongseob Oh  1 , Jong Hyuk Yoon, Soonyong Park, Yung Joon Yoo
Affiliations

Simultaneous quantification of total and conjugated ubiquitin levels in a single immunoblot

Choongseob Oh et al. Anal Biochem. .

Abstract

Ubiquitin (Ub) is a posttranslational modifier, and total Ub (UbT) is always in dynamic equilibrium among free Ub (UbF), activated Ub (UbA), and conjugated Ub (UbC) in the forms of mono-Ub, thioester-bond-linked Ub, and peptide-bond-linked Ub, respectively. In this study, we developed a simple method to simultaneously determine the levels of UbT, UbF+UbA, and UbC in a single immunoblot and demonstrated its reliability and reproducibility by determining [UbT], [UbF+UbA], and [UbC] in various mouse tissues and cultured cells.

Keywords: Ubiquitin; Ubiquitination; Western blot.

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