DETECTION METHOD OF INTRACELLULAR UBIQUITINATION

摘要

PROBLEM TO BE SOLVED: To provide a method for quantitatively detecting polyubiquitination and/or deubiquitination of a target protein in a cell.

SOLUTION: The method for detecting polyubiquitination and/or deubiquitination of a target protein in a cell includes the following steps of: (1) preparing a transformed cell, which expresses both of a fusion protein (X) of a target protein (A) and a tag (T1) and a fusion protein (Y) of ubiquitin (B) and a tag (T2), in which the fusion protein (X) is labeled with a fluorescent pigment (D1) and the fusion protein (Y) is labeled with a fluorescent pigment (D2); (2) incubating the transformed cell under the conditions to promote polyubiquitination and/or deubiquitination of the target proteins; (3) irradiating the transformed cell with laser light; and (4) measuring a fluorescent life of fluorescence emitted by the transformed cell.

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