Product Pathways - Cytoskeletal Signaling
Phospho-Myosin Light Chain 2 (Ser19) Mouse mAb #3675
PhosphoSitePlus® protein, site, and accession data: MRLC1
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IF-IC | H M R B Pg | Endogenous | 18 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
B=Bovine
Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Phospho-Myosin Light Chain 2 (Ser19) Mouse mAb detects endogenous levels of myosin light chain 2 (smooth muscle) only when phosphorylated at serine 19. This antibody does not cross-react with the cardiac isoform of myosin light chain 2.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser19 of human myosine light chain 2 (smooth muscle).
Western Blotting
Western blot analysis of extracts from HEK293, C2C12, and C6 cells, untreated (-) or lamda phosphatase treated (+), using Myosin Light Chain 2 Antibody #3672 (upper) or Phospho-Myosin Light Chain 2 (Ser19) Mouse mAb (lower).
IF-IC
Confocal immunofluorescent images of HeLa cells, untreated (left) or phosphatase-treated (right), labeled with Phospho-Myosin Light Chain 2 (Ser19) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Background
Myosin is composed of six polypeptide chains: two identical heavy chains and two pairs of light chains. Myosin light chain 2 (MLC2), also known as myosin regulatory light chain (MRLC or RLC, LC20), has many isoforms depending on its distribution. In smooth muscle, MLC2 is phosphorylated at Thr18 and Ser19 by myosin light chain kinase (MLCK) in a Ca2+/calmodulin-dependent manner (1). This phosphorylation is correlated with myosin ATPase activity and smooth muscle contraction (2). ROCK also phosphorylates Ser19 of smooth muscle MLC2, which regulates the assembly of stress fibers (3). Phosphorylation of smooth muscle MLC2 at Ser1/Ser2 and Ser9 by PKC and cdc2 has been reported to inhibit myosin ATPase activity (4,5). Phosphorylation by cdc2 controls the timing of cytokinesis (5). Transgenic mice lacking phosphorylation sites on the cardiac muscle isoform show morphological and functional abnormalities (6).
- Ikebe, M. and Hartshorne, D.J. (1985) J. Biol. Chem. 260, 10027-10031.
- Tan, J. L. et al. (1992) Annu. Rev. Biochem. 61, 721-759.
- Totsukawa, G. et al. (2000) J. Cell Biol. 150, 797-806.
- Ikebe, M. et al. (2000) J. Biol. Chem. 262, 9569-9573.
- Satterwhite, L. L. et al. (1992) J. Cell Biol. 118, 595-605.
- Sanbe, A. et al. (1999) J. Biol. Chem. 274, 21085-21094.
Application References
- Sakurada, K. et al. (1998) Dynamics of myosin light chain phosphorylation at Ser19 and Thr18/Ser19 in smooth muscle cells in culture. Am. J. Physiol. 274, 1563-1572. Applications: IC-IF Western Blotting
- Totsukawa, F. et al. (2000) Distinct roles of ROCK (Rho-kinase) and MLCK in spatial regulation of MLC phosphorylation for assembly of stress fibers and focal adhesions in 3T3 fibroblasts. J. Cell Biol. 150, 797-806. Applications: IC-IF
- Watanabe, T. et al. (2007) Mol Biol Cell 18, 605-16. Applications: Western Blotting
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Companion Products
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 3671 Phospho-Myosin Light Chain 2 (Ser19) Antibody
- 3672 Myosin Light Chain 2 Antibody
- 3674 Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
