ADP-Glo Kinase Assay + MLCK Kinase Enzyme System
Recombinant human MLCK (1425-1776) was expressed by baculovirus in Sf9 insect cells using a N-terminal GST tag. MLCK or myosin light chain kinase is a muscle member of the immunoglobulin gene superfamily and is a calcium/calmodulin-dependent protein kinase. MLCK is a regulatory protein for smooth muscle contraction, which acts by phosphorylating 20-kDa myosin light chain (MLC20) to activate the myosin ATPase activity. The kinase activity for the phosphorylation is localized at the central part of MLCK, which is also furnished with actin-binding activity at its N terminal and myosin-binding activity at its C terminal. Phosphorylation of regulatory light chain of myosin plays an important role in controlling the morphological changes seen during cell division. 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: MLCK, 10microg (Human, recombinant; amino acids 1425-1776). MW: ~70kDa. Substrate: MRCL3 Peptide (KKRPQRATSN-VFAM-NH2); derived from human myosin regulatory light chain MRCL3 (amino acid 11-24). Other: Reaction Buffer, DTT, Ca2+/Calmodulin Solution. MLCK NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/91807/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.