Product Description
MLCK (Myosin light chain kinase), also known as MYLK or MLCK1, is a member of the serine/threonine protein kinase family (myosin light chain kinase subfamily) that plays a pivotal role in smooth muscle contraction, cytoskeletal dynamics, cell migration, and vascular permeability, as well as in epithelial barrier function and non‑muscle cell motility. MLCK is activated by various stimuli including calcium/calmodulin binding, phosphorylation by Rho‑associated kinases (ROCK), and inflammatory mediators such as TNFα, leading to phosphorylation of myosin regulatory light chain (MLC), thereby promoting actin‑myosin cross‑bridging and contraction. Beyond its role in normal vascular homeostasis and tissue barrier integrity, dysregulation of MLCK signaling—through overexpression, alternative splicing variants, or genetic polymorphisms—is closely associated with inflammatory bowel disease (such as ulcerative colitis), asthma, acute lung injury, vascular disorders (e.g., hypertension, atherosclerosis), and multiple cancers (including colorectal cancer and breast cancer metastasis), making it a highly validated target for anti‑inflammatory therapy, vascular disease treatment, and cancer drug discovery.
Screeningbio’s MLCK/MYLK Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying MLCK/MYLK kinase activity and evaluating the potency of tyrosine kinase inhibitors (TKIs).
This kit utilizes the ADP-Glo Kinase Assay platform, a luminescent technology that measures the amount of ADP produced during the kinase reaction.
The assay is performed in two distinct steps:
Kinase Reaction: MLCK/MYLK catalyzes the transfer of a phosphate group from ATP to a specific substrate, resulting in the production of phosphorylated substrate and ADP.
ADP-Glo™ Reagent Addition: After the kinase reaction, the ADP-Glo™ Reagent is added to terminate the reaction and deplete any remaining unreacted ATP.
Kinase Detection: The Kinase Detection Reagent is added to simultaneously convert the produced ADP back into ATP, which is then used by luciferase to generate a luminescent signal.
The light intensity produced is directly proportional to the ADP concentration and, consequently, to the enzymatic activity of the MLCK/MYLK kinase.
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