NanoBRET Intracellular E3 Ligase Detection Reagents, VHL

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The PRKG1 Kinase Enzyme System contains:

• PRKG1 Kinase, 10μg (Human, recombinant full-length). MW: ~100kDa.
• RSK Substrate(KRRRLSSLRA); derived from human 40S ribosomal protein S6 (amino acid 230239).
• cGMP Solution + Reaction Buffer A + DTT.

Recombinant full-length human PRKG1 (PKG) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. PRKG1 is a homodimer, with each monomer containing a regulatory cGMP-binding domain and a catalytic domain. PRKG1 was shown to be expressed at highest levels in bladder, uterus, adrenal gland and fallopian tube. PRKG1 plays an important stimulatory role in platelet activation. Expression of recombinant PRKG1 in a reconstituted cell model enhanced von Willebrand factor-induced activation of the platelet integrin alpha-IIb/beta-3. PRKG1 knockout mice showed impaired platelet responses to VWF or low doses of thrombin and prolonged bleeding time. Human platelet aggregation induced by VWF or low-dose thrombin was inhibited by PRKG1 inhibitors but enhanced by cGMP.

PRKG1 NCBI Database Entry.

The PRKG1 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.