Patent ID: 6348310
Filing Date: 2002-02-19
Classification: C07K,C12Q

Abstract:
A method of determining the presence or activity of a selected protein kinase, comprising:a. conjugating a biotinylating reagent to a peptide substrate forming a biotinylated peptide substrate containing a biotin moiety; wherein the biotin moiety does not result in a steric hindrance of the biotinylated peptide substrate accessibility to the protein kinase; b. adding a sufficient quantity of the biotinylated peptide substrate to a solution containing the selected protein kinase; c. incubating the selected protein kinase with the biotinylated peptide substrate under conditions where the selected protein kinase is active for a time sufficient to form a biotinylated peptide product; d. binding the biotinylated peptide product to a biotin-binding matrix, wherein the matrix has a sufficient binding capacity and specificity for the biotinylated peptide product; and e. determining the presence of the activity of the selected protein kinase.