The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The RET Kinase Enzyme System contains:
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RET Kinase, 10μg (Human, recombinant; amino acids 658–end). MW: ~74kDa.
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IGF1Rtide (KKKSPGEYVNIEFG) Substrate; derived from human IRS-1 protein residues 891–902.
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Reaction Buffer, DTT.
Recombinant human RET (amino acids 658–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. RET gene codes for a transmembrane tyrosine kinase, which is a subunit of a multimeric complex that acts as a receptor for four structurally related molecules: GDNF, neurturin, artemin and persephin. Germline mutations of RET cause a dominantly inherited dysgenesis of the enteric nervous system known as Hirschsprung's disease. RET is constitutively activated by point mutations in hereditary medullary thyroid carcinomas (MTCs). Several single nucleotide polymorphisms of the RET gene have been described. Multiple endocrine neoplasia type 2A (MEN 2A) have been reported to be associated with two mutations of the proto-oncogene RET.
RET NCBI Database Entry.
The RET 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.