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Technical Data Sheet

MM3441

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Myosin Light Chain (MLC20)
Mouse Monoclonal IgM
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms, Ck
20 kDa

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Western blot analysis of C2C12 cells untreated (lanes 1, 3, 5, & 7) or treated with Lambda phosphatase (lanes 2, 4, 6, & 8). The blots were probed with monoclonal anti-MLC20 (clone MY-21) (lanes 1 & 2), polyclonal anti-MLC (Ser-19) phosho-specific (lanes 3 & 4), anti-MLC (Ser-1) phosho-specific (lanes 5 & 6), and anti-MLC (a.a. 11-22) (lanes 7 & 8) .

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Immunocytochemical labeling of phosphorylated MLC in paraformaldehyde fixed A7r5 cells. The cells were dual-labeled with anti-MLC (MM3441; middle) and anti-MLC (MP4201; top left), anti-MLC (Ser-19) (MP4221; middle left) and anti-MLC (Ser-1) (MP3461; bottom left). Goat anti-Mouse DyLight® 488 and Goat anti-Rabbit DyLight® 594 were used for detection of primary antibodies. The overlay of staining patterns are shown to the right.

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Application
Dilution


ELISA
1:1000


ICC
1:250


WB
1:250



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Both smooth muscle and nonmuscle myosin II activity is regulated by the phosphorylation state of the myosin regulatory light chain (MLC, MRLC, MLC20, Myl9). Phosphorylation of MLC at Thr-18 and Ser-19 activates myosin II motor activity and increases myosin filament stability. This activation has important roles in various cell motile processes. By contrast, other phosphorylation sites on MLC may inhibit myosin II activity. PKC phosphorylates Ser-1/Ser-2 and Thr-9 in MLC, and this phosphorylation decreases activated myosin II interaction with actin, as well as inhibits MLC interaction with the activation site kinase, myosin light-chain kinase. The Ser-1/Ser-2 region may be the major inhibitory site since Ser-1 is phosphorylated during PDGF-induced stress fiber disassembly and expression of unphosphorylatable MLC20 at the Ser-1/Ser-2 site suppresses this disassembly. Thus, inhibition of myosin II activity through phosphorylation of Ser-1/Ser-2 may have important roles in growth factor-induced reorganization of actomyosin filaments.


Background References
Sellers, J.R. (1991) Curr. Opin. Cell Biol. 3:98.
Tan, J.L. et al. (1992) Annu. Rev. Biochem. 61:721.
Komatsu, S. & Ikebe, M. (2007) Mol. Biol. Cell 18:5081.
Immunogen
Clone MY-21 was generated from myosin light chains in chicken lens membrane extracts. The antibody reacts with myosin from chicken, pig, bovine, rabbit, and human muscle.
Buffer and Storage
Mouse monoclonal is provided as ascites fluid with 0.02% sodium azide. Store at 4°C, stable for 6 months. For long term storage, aliquot and store at -20°C.
Specificity
The antibody detects a 20 kDa* band corresponding to the molecular weight of myosin light chain in human A431, mouse C2C12, and rabbit fibroblasts.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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MP3461
Myosin Light Chain (Ser-1), phospho-specific Rabbit Polyclonal

MP4201
Myosin Light Chain (a.a. 11-22) Rabbit Polyclonal

MP4221
Myosin Light Chain (Ser-19), phospho-specific Rabbit Polyclonal

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