ADP-Glo Kinase Assay + ULK1 Kinase Enzyme System
Recombinant human ULK1 (1-649) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ULK1 is a serine/threonine protein kinase that plays a critical role during the initial stages of autophagy, which is a vital response to nutrient starvation. The conserved C-terminal domain (CTD) of ULK1 controls the regulatory function and localization of the protein. Knockdown of ULK1 inhibits the autophagic response as well as inhibiting rapamycin-induced autophagy consistent with a role downstream of mTOR. ULK1 forms a complex with FIP200 and ATG13, and this complex is essential for starvation-induced autophagy. Both FIP200 and ATG13 are critical for correct localization of ULK1 to the pre-autophagosome and stability of ULK1 protein. ULK1 is phosphorylated by the mTOR pathway in a nutrient starvation-regulated manner. 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 chemical compounds have 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. Kinase Enzyme System contains: Kinase: ULK1, 10ug (Human, recombinant; amino acids 1-649). MW: ~125kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT. ULK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/8408/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.