P27 ubiquitination assay and methods of use

Details P27 ubiquitination assay and methods of use P27 ubiquitination assay and methods of use

G - Physics

01

N

G01N 21/75 (2006.01) G01N 21/76 (2006.01) G01N 33/53 (2006.01) G01N 33/532 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2584131

This invention relates to easy and reliable assays for the ubiquitination of p27. Because the assay can be performed as a plate capture assay, or as a homogenous time-resolved fluorescence resonance energy transfer assay, with defined, easily replicatable reaction conditions, it is particularly useful for high-throughput screening of, for example, a prospective anticancer agent. The invention provides methods to determine the amount of ubiquitination of p27, or of any protein, and use of the method to identify compounds that modulate the ubiquitination of p27, or any protein.

L'invention concerne des analyses faciles et fiables de l'ubiquitination de p27. Etant donné que l'analyse peut être mise en oeuvre sous forme d'analyse par capture sur plaque, ou sous forme d'analyse de transfert d'énergie de résonance de fluorescence en temps résolu, homogène, dans des conditions de réactions définies, pouvant facilement être répétées. L'invention est particulièrement utile pour le criblage à haut rendement de, par exemple, un agent anticancéreux éventuel. L'invention concerne des méthodes permettant de déterminer la quantité d'ubiquitination de p27, ou de n'importe quelle protéine, ainsi que l'utilisation de la méthode pour identifier des composés qui modulent l'ubiquitination de p27, ou de n'importe quelle protéine.

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