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SignaTECT DNA-Dependent Protein Kinase Assay System

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Convenient, 1-Hour Kinase Activity Assays

    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 [?-32P]ATP.

    Method Overview

    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.

    Advantages of the SAM2® Membranes

    • High binding capacity and high specificity characteristics produce lower backgrounds and higher signal-to-noise ratios compared to the traditional P81 phosphocellulose method of capture and measurement.
    • The perforated and numbered membrane allows researchers to measure from 1 up to 96 kinase reactions and does not require as much "hands-on" manipulation as other methods used to measure kinase activity.
    • Can be used under a variety of buffer/reaction conditions (e.g., cell extracts), which many other methods do not allow.
    • High binding capacity allows use of the SignaTECT® Systems for kinetic studies.
  • Protocols

    Complete Protocol

    Download PDF

    SignaTECT® Protein Kinase C (PKC) Assay System Technical Bulletin

    PDF (519 KB)

    Download PDF

    SignaTECT® Protein Tyrosine Kinase Assay System Technical Bulletin

    PDF (792 KB)

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    SignaTECT® Calcium/Calmodulin-Dependent Protein Kinase Assay System Technical Bulletin

    PDF (551 KB)

    Download PDF

    SignaTECT® DNA-Dependent Protein Kinase Assay System Technical Bulletin

    PDF (591 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources