MARK3 Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The MARK3 Kinase Enzyme System contains:

• MARK3 Kinase, 10μg (Human, full-length recombinant). MW: ~110kDa
• CHKtide Substrate (1mg/ml), (KKKVSRSGLYRSPSMPENLNRPR); derived from human CDC25C protein isoform A (amino acid 205–225)
• 5X Reaction Buffer, 0.1M DTT

Recombinant full-length human MARK3 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Microtubule affinity-regulating kinase 3 (MARK3) is a member of the PAR-1/MARK kinase family, which play critical roles in polarity and cell cycle control and are regulated by 14-3-3 scaffolding proteins, as well as the LKB1 tumour suppressor kinase and atypical protein kinase C (PKC). MARK3 is a dual-specificity protein kinase that controls entry into mitosis by dephosphorylating CDC2. MARK3 seems to be a positive regulator of the beta-catenin pathway and an inhibitor of the JNK pathway. MARK3, a regulator of polarity, is also a modulator of Wnt-beta-catenin signalling, indicating a link between two important developmental pathways.

Application Note

NCBI Database Entry

The MARK3 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.