Patent ID: 6066462
Filing Date: 2000-05-23
Classification: C07K,C12Q

Abstract:
A method of determining the presence or activity of a selected protein kinase, comprising:a. providing a biotinylated peptide substrate containing a biotin moiety, wherein the biotin moiety is conjugated to the peptide substrate at its N-terminal amino group via a linker at least six carbon atoms in length;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 phosphorylated, 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 to capture essentially all of the biotinylated peptide substrate and biotinylated peptide product from solution; ande. detecting the presence and measuring the activity of the selected protein kinase.