ADP-Glo Kinase Assay + PKC mu Kinase Enzyme System
Description
- Overview
- Protocols
- Specifications
- Resources
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 PKCu Kinase Enzyme System contains:
- PKCu Kinase, 10ug (Human, recombinant full-length). MW: ~131kDa.
- CREBtide (KRREILSRRPSYR) Substrate; derived from human CREB1 isoform A (amino acids 109–121).
- Reaction Buffer, DTT.
Recombinant full-length human PKCu was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Protein Kinase C mu is a member of the protein kinase C (PKC) family that differs from the other PKC isoenzymes in structural and enzymatic properties. PKCu is ubiquitous in nature with the highest expression in the thymus, lung and peripheral blood mononuclear cells. PKCu forms a complex in vivo with a phosphatidylinositol 4-kinase and a phosphatidylinositol-4-phosphate 5-kinase. A region of PKCu between the amino-terminal transmembrane domain and the pleckstrin homology domain is shown to be involved in the association with the lipid kinases.
The PKCu 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.
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Protocols
Complete Protocol
PKC mu Kinase Enzyme Protocol
PDF (914 KB)
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Certificate of Analysis
Lookup Certificate of AnalysisStorage Conditions
LESS THAN -65C
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
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Resources
Posters
- Bringing Assays and Automation Together for Lower Throughput Kinase Profiling Poster
- Automation of ADP-Glo Universal Kinase Assay For HTS Screening AND Profiling Applications Poster
Other
