Methods for determination of protein phosphatase activity,...

Details Methods for determination of protein phosphatase activity,... Methods for determination of protein phosphatase activity,...

C - Chemistry, Metallurgy

12

Q

C12Q 1/48 (2006.01) C12N 9/12 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2694329

One aspect of the present disclosure encompasses methods for determining a protein kinase or phosphatase activity in a biological sample, comprising: contacting in a reaction mix a first test sample and a fluorescently-labeled peptide substrate capable of being modified by a protein phosphatase or a protein kinase, contacting the reaction mix with a TiO2 matrix, thereby partitioning fluorescently-labeled phosphorylated peptide from fluorescently- labeled dephosphorylated peptide; and determining the fluorescence of the fluorescently-labeled dephosphorylated peptide, thereby determining a protein kinase or phosphatase activity.

L'invention concerne des procédés de détermination d'une activité de protéine kinase ou phosphatase dans un échantillon biologique. Le procédé consiste à mettre en contact, dans un mélange de réaction, un premier échantillon d'essai et un substrat de peptide marqué de manière fluorescente apte à être modifié par une protéine phosphatase ou une protéine kinase ; à mettre en contact le mélange de réaction avec une matrice de TiO2, séparant ainsi le peptide phosphorylé marqué de manière fluorescente du peptide déphosphorylé marqué de manière fluorescente ; et à déterminer la fluorescence du peptide déphosphorylé marqué de manière fluorescente et donc une activité de protéine kinase ou phosphatase.

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