ADP-Glo Kinase Assay + TAK1-TAB1 Kinase Enzyme System
Recombinant human proteins TAK1 (1-303) and TAB1 (437-end), linked by a small peptide (DFGGGGG), were expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. TAK1 is a serine/threonine protein kinase that mediates signaling by TGF beta and morphogenetic protein (BMP) (1). In response to IL-1, TAK1 forms a kinase complex with TAB1, and this complex is required for the activation of nuclear factor kappa B (NF kappa B). TAK1 also can activate MAPK8/JNK and MAP2K4/MKK4 and thus play a role in the cell response to environmental stress. TAK1 is essential for thymocyte development and activation, and deletion of TAK1 prevents maturation of single-positive thymocytes displaying CD4 or CD8 (2). Thymocytes lacking TAK1 fail to activate NF kappa B and JNK and are prone to apoptosis upon stimulation. 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: TAK1-TAB1, 10ug (Human, recombinant; TAK1 (1-303) and TAB1 (437-end)). MW: ~74kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT. TAK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/6885/. TAB1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/10454/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.