Method for quantifying phosphokinase activity on proteins

Details Method for quantifying phosphokinase activity on proteins Method for quantifying phosphokinase activity on proteins

C - Chemistry, Metallurgy

07

K

C07K 16/28 (2006.01) C07K 16/18 (2006.01) C12Q 1/48 (2006.01) G01N 33/53 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2423979

The invention involves a method for measuring phosphorylation of proteins and, as such, is an indicator of protein kinase activity. The method involves the in vitro phosphorylation of a target protein but subjecting that protein (non- phosphorylated) to reaction mixture containing all reagents, including phosphokinase which allow the creation of a phosphorylated form of protein. The phostphorylated protein is measured by contacting it with an antibody specific for the phosphorylation sites(s). The invention includes antibodies useful in practicing the methods of the invention. The invention particularly relates to phosphorylation of Tau, Rb and EGFR proteins and antibodies specific for the site of phosphorylation of the Tau, Rb or EGFR proteins.

L'invention porte sur un procédé de mesure de la phosphorylation de protéine servant d'indicateur de l'activité de protéine kinases. Le procédé comporte la phosphorylation in vitro d'une protéine cible, obtenu en soumettant ladite protéine (non-phosphorylée) à un mélange contenant tous les réactifs dont la phosphokinase, pour donner la forme phosphorylée de la protéine. On mesure la protéine phosphorylée en la mettant en contact avec un anticorps spécifique du ou des sites de phosphorylation. L'invention porte également sur les anticorps utiles pour mettre en oeuvre lesdits procédé, et en particulier sur la phosphorylation des protéines Tau, Rb et EGFR, et sur les anticorps spécifiques des site de phosphorylation des protéines Tau, Rb ou EGFR.

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