The SignaTECT® Protein Kinase Assay Systems provide a rapid, radioactivity-based method for detection of kinase activity. The method is based on capture of phosphorylated labeled substrates on the SAM2® Biotin Capture Membrane, which offers low background and high signal-to-noise ratios, even for complex samples such as crude cell extracts, while retaining the high substrate capacity necessary to maintain optimum reaction kinetics.
Each SignaTECT® System contains a highly specific biotinylated peptide substrate for the appropriate kinase as well as the necessary reaction components. The researcher must supply [γ-32P]ATP.
After the kinase reaction is complete, the biotinylated, 32P-labeled substrate is recovered from the reaction mix using the SAM2® Biotin Capture Membrane. After samples are spotted onto the SAM2® Membrane a series of short washes are performed to remove nonspecific label. The process is complete in less than 1 hour. The SAM2® Membrane is prenumbered and partially cut so that individual squares can be easily identified, separated and placed in scintillation vials. Alternatively, the intact SAM2® Membrane can be analyzed by phosphorimaging or conventional autoradiography.