Anti-MLCK Antibody, clone 5K6 ZooMAb^® Rabbit Monoclonal

Myosin light chain kinase, smooth muscle (UniProt: Q15746; also known as MLCK, smMLCK, Kinase-related protein, KRP, Telokin, EC:2.7.11.18) is encoded by the MYLK (also known as MLCK, MLCK1, MYLK1) gene (Gene ID: 4638) in human. MYLK gene is mapped to the human chromosome location 3q21.1. This gene encodes telokin, non-muscle and the smooth muscle isoforms of MLCK. The encoded protein MLCK is a Ca2+/calmodulin dependent myosin light chain phosphorylating agent. This enzyme plays a pivotal role in the phosphorylation of the regulatory light chains of myosin which are essential for the shortening and tension development of smooth muscle cells resulting in smooth muscle contraction. MLCK also assists the interaction of myosin with actin. It is expressed in a wide variety of adult and fetal tissues especially in skeletal, cardiac, smooth and mammalian non-muscle cells. MLCK localizes predominantly in the cytoplasm and at the perijunctional actomyosin ring in epithelial cells, a localization that is enhanced under inflammatory conditions. Structurally, MLCK is characterized by multiple Ig-like C2-type domains, Fibronectin type-III domain, a protein kinase domain and several tandem repeat regions. Multiple isoforms of MLCK have been described that are produced by alternative promoter usage and alternative splicing. Non-muscle isoform 2 is the dominant splice variant expressed in various tissues, which is essential for maintaining endothelial barrier integrity and responding to inflammatory stimuli. Loss of function of the protein has been linked to megacystis microcolon intestinal hypoperistalsis syndrome. Notably, genetic variants in MLCK, such as single nucleotide polymorphisms (SNPs), have been associated with altered promoter activity and increased susceptibility to inflammatory diseases. Recent studies indicate that mis-localization of MLCK can disrupt endothelial barrier function, contributing to pathologies like asthma and ARDS. This ZooMAb^® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Sun, X., et al. (2021). Clin Sci. 135(7); 963-977; Wang, T., et al. (2018). Pulm Circ. 8(2); 2045894018764171).

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

KLH-conjugated linear peptide corresponding to 25 amino acids from the internal region of human MLCK.

Quality Control Testing

Evaluated by Western Blotting in rat colon tissue lysate.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected MLCK in rat colon tissue lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected MLCK in lysate from human colon tissue.

Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected MLCK in HepG2 cells.

Affinity Binding Assay: A representative lot of this antibody bound MLCK peptide with at least one hundred thousand-fold (100,000X) higher affinity than with non-specific control peptide.

Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected MLCK in human uterus tissue sections.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 5K6 is a ZooMAb^® recombinant rabbit monoclonal antibody that specifically detects MLCK. It targets an epitope within 25 amino acids from the internal region.

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.