ADP-Glo Kinase Assay + PAK1/CDC42 Kinase Enzyme System

by Promega
Available SKUs: V4479 |
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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 PAK1/CDC42 Kinase Enzyme System contains:

    • PAK1/CDC42 Kinase, 10ug (Human, recombinant full-length). MW: ~96kDa (PAK1).
    • PAKtide (RRRLSFAEPG) Substrate.
    • Reaction Buffer, DTT, GTP Solution.

    Recombinant full-length human PAK1 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag and combined with CDC42 at a ratio of 1:4 (PAK1:CDC42) in vitro. PAK1 is a member of the p21-activated kinases (PAKs), which have been implicated in the regulation of cell morphology, motility and transformation. These serine/threonine kinases are activated by and are effectors of small GTPases, CDC42 and RAC. PAK1 is one of the Group I PAKs, which also include PAK2 and PAK3. PAK1 is a key regulator of the actin cytoskeleton, adhesion and cell motility. Inactive dimeric PAK1 is mainly cytosolic and interaction with the activators Cdc42-GTP and Rac1-GTP stimulates the kinase at the sites of cellular protrusions forming adhesions to the extracellular matrix.

    PAK1/CDC42 NCBI Database Entry.

    The PAK1/CDC42 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

    PAK1/CDC42 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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