ADP-Glo Kinase Assay + TOPK Kinase Enzyme System

by Promega
Available SKUs: V4095 |
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Description

  • Overview
  • Protocols
  • Specifications
  • Resources
  • Kinase-background-v2light

    Convenient, Scalable Kinase Profiling

    The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The TOPK Kinase Enzyme System contains:

    • TOPK Kinase, 10ug (Human, recombinant full-length). MW: ~68kDa.
    • Native Swine Myelin Basic Protein (MBP) Substrate.
    • Reaction Buffer, DTT.

    Recombinant full-length human TOPK was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. TOPK is a MAPK kinase that phosphorylates p38 MAPK and is activated in a cell-cycle-dependent manner in neuronal progenitor cells in vitro. Expression of TOPK is detected in male germ line progenitor cells, activated T-cells and a variety of lymphomas and leukemias. In vitro studies have shown that activated TOPK phosphorylates p38 MAPK but not JNK or ERK. TOPK activation requires phosphorylation by both the M-phase CDK1/CyclinB kinase complex and another unknown kinase, possibly Raf C or Raf A. TOPK may play an important role in linking extracellular signals to an intracellular state, possibly allowing extracellular influence on the cell-cycle-related processes of proliferation or differentiation.

    TOPK NCBI Database Entry.

    The TOPK Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.

    Use with ADP-Glo™ Kinase Assay

    The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.

     

    See all Kinase Enzyme Systems available from Promega.

  • Protocols

    Complete Protocol

    Download PDF

    TOPK Kinase Enzyme Protocol

    PDF (925 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    LESS THAN -65C

    Use Restrictions

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

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