M9850-07
Clone Type
PolyclonalHost
RabbitSource
HumanIsotype
IgGGrade
Affinity PurifiedApplications
WBCrossreactivity
Bo Hu Mo RbShipping Temp
Blue IceStorage Temp
-20°CRabbit Anti-Myosin Light Chain Kinase, Smooth Muscle, phosphorylated (Ser1760) (MLCK)
Smooth Muscle Myosin Light Chain Kinase (MLCK) is a multifunctional regulatory protein of smooth muscle contraction (SMC) and a key element in ligand-mediated endothelial cell gap formation and vascular permeability, motility and morphology. Smooth muscle MLCK exists in at least two isoforms, short (~150kD) and long (210kD) which are identical except for an extended amino terminus with two additional putative actin-binding motifs in the long isoform. MLCK is phosphorylated by several kinases including protein kinase A (PKA) and mediates its function by phosphorylating 20kD myosin light chain (MLC20). Phosphorylation of MLCK inhibits the actin-activated ATPase of myosin II by reducing its affinity for actin. MLCK possesses a counter-balancing role in vascular regulation, by mediating vasoconstriction via direct action on SMCs and vasodilation via action on endothelial cells (ECs).
Applications
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilutions
Western Blot: 1:1000 Optimal dilutions to be determined by the researcher.
Positive Controls
KCl-treated rabbit muscle extracts; COS cells over-expressing rabbit smooth muscle short MLCK, phosphorylated in vitro by PKA.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic phosphopeptide derived from the region of human myosin light chain kinase (MLCK) that contains serine 1760.
Form
Supplied as a liquid in Dulbecco’s PBS (without Mg2+ and Ca2+), pH 7.3, 1mg/ml BSA (IgG, protease free), 0.05% sodium azide, 50% glycerol.
Purity
Purified by immunoaffinity chromatography.
Specificity
Recognizes MLCK when phosphorylated at Ser1760. Species Crossreactivity: rabbit, mouse and bovine. Species Sequence Homology: human,100%.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
References
1. Isotani, E., et al. (2004) Real-time evaluation of myosin light chain kinase activation in smooth muscle tissues from a transgenic calmodulin-biosensor mouse. Proc. Nat’l. Acad. Sci. USA 101(16):6279-6284. 2. Dulyaninova, N.G. and A.R. Bresnick (2004) The long myosin light chain kinase is differentially phosphorylated during interphase and mitosis. Exp. Cell Res. 299(2):303-314. 3. Totsukawa, G., et al. (2004) Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts. J. Cell. Biol. 164(3):427-439. 4. Tinsley, J.H., et al. (2004) Isoform-specific knockout of endothelial myosin light chain kinase: closing the gap on inflammatory lung disease. Trends Pharmacol. Sci. 25(2):64-66. 5. Kamm, K.E. and J.T. Stull (2001) Dedicated myosin light chain kinases with diverse cellular functions. J. Biol. Chem. 276(7):4527-4530. 6. Takaishi, K., et al. (2000) Localization and activity of myosin light chain kinase isoforms during the cell cycle. J. Cell. Biol. 151(3):697-708. 7. Watanabe, H., et al. (2001) Myosin light-chain kinase regulates endothelial calcium entry and endothelium-dependent vasodilation. FASEB J. 15(2):282-284. 8. Smith, L., et al. (2002) Properties of long myosin light chain kinase binding to F-actin in vitro and in vivo. J. Biol. Chem. 277(38):35597-35604.USBio References
No references available