Assay for kinases and phosphatases

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/00 (2006.01) C12Q 1/42 (2006.01) C12Q 1/44 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2410823

Disclosed is a method and corresponsind kit for assaying the presence, activity, or both, of an enzyme classified within an enzyme classification selected from the group consisting of EC 2.7.1, EC 3.1.3, and EC 3.1.4. The method generally includes the steps of reacting an enzyme with a substrate for a time sufficient to yield phosphorylated or dephosphorylated product; contacting the product with a binding matrix, whereby product is adhered to the matrix; and then analyzing the matrix for presence of, amount of, or both the presence and the amount of the product fixed to the matrix, whereby the presence, the activity, or both the presence and activity of the enzyme can be determined.

L'invention concerne un procédé et un kit correspondant destinés à faire des dosages pour détecter la présence et/ou l'activité d'une enzyme qui entre dans la classification des enzymes appartenant au groupe constitué de EC 2.7.1, EC 3.1.3 et EC 3.1.4. De façon générale, le procédé consiste à faire réagir une enzyme avec un substrat pendant un laps de temps suffisant pour obtenir un produit phosphorylé ou déphosphorylé; mettre le produit en contact avec une matrice de liaison, le produit se collant ainsi à la matrice; et analyser ensuite la matrice pour détecter la présence et/ou la quantité de produit fixé à la matrice. La présence et/ou l'activité de l'enzyme peut ainsi être déterminée.

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