ADP-Glo Kinase Assay + MAPKAPK5 Kinase Enzyme System
Recombinant full-length human MAPKAPK5 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. MAPKAPK5 is a member of the serine/threonine kinase family that responds to cellular stress and proinflammatory cytokines. MAPKAPK5 is activated through its phosphorylation by MAP kinases including MAPK1/ERK, MAPK14/p38-alpha and MAPK11/p38-beta (1). MAPKAPK5 is activated in HeLa cells in response to cellular stress and proinflammatory cytokines. MAPKAPK5 activity is regulated by p38-alpha and p38-beta both in vitro and in vivo, and thr-182 is the regulatory phosphorylation site of MAPKAPK5 (2). In vitro, MAPKAPK5 kinase phosphorylates heat shock protein HSP27 at its physiologically relevant sites. 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: MAPKAPK5, 10ug (Human, recombinant full-length). MW: ~79kDa. Substrate: HSP27tide (RRLNRQLSVA-amide); derived from the mouse HSP27 (amino acids 80-85). Other: Reaction Buffer, DTT. MAPKAPK5 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/8550/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.