Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Myosin Light Chain 2 Antibody Sampler Kit #9776

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Myosin Light Chain 2 (Ser19) Antibody #3671 40 µl W IF-IC H M R Dm (C) (X) (Z) (B) (Pg) 18 Rabbit
Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody #3674 40 µl W IF-IC H M (R) (C) (X) (Z) (B) (Pg) 18 Rabbit
Myosin Light Chain 2 Antibody #3672 40 µl W H M R (C) (B) (Pg) 18 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  C=Chicken  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  B=Bovine  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

The antibodies in the Myosin Light Chain 2 Antibody Sampler Kit detect endogenous levels of total myosin light chain 2 (smooth muscle), Myosin Light Chain 2 when phosphorylated at Ser19, or when dually phosphorylated at Thr18 and Ser19, respectively. The antibodies do not cross-react with the cardiac isoform of Myosin Light Chain 2 and the Myosin Light Chain 2 Antibody does not cross-react with Mysoin Essential Light Chain.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells stimulated with ionophore A23187 for the indicated times, using Phospho-Myosin Light Chain 2 (Ser19) Antibody #3671.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, PL45, A431, C2C12 and C6 cells using Myosin Light Chain 2 Antibody #3672.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells stimulated with ionophore A23187 for the indicated times using Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody #3674.


Description

The Myosin Light Chain 2 Antibody Sampler Kit provides an economical means to detect total, phosphorylated, and dual-phosphorylated myosin light chain 2. The kit contains enough primary and secondary antibody to perform four western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Myosin Light Chain 2 (smooth muscle isoform), a synthetic phosphopeptide corresponding to residues surrounding Ser19 of human myosin light chain 2, or to residues surrounding Thr18/Ser19 of human myosin light chain 2, respectively. Antibodies are purified by protein A and peptide affinity chromatography.

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), RLC, or 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).

  1. Ikebe, M. and Hartshorne, D.J. (1985) J. Biol. Chem. 260, 10027-10031.
  2. Tan, J. L. et al. (1992) Annu. Rev. Biochem. 61, 721-759.
  3. Totsukawa, G. et al. (2000) J. Cell Biol. 150, 797-806.
  4. Ikebe, M. et al. (2000) J. Biol. Chem. 262, 9569-9573.
  5. Satterwhite, L. L. et al. (1992) J. Cell Biol. 118, 595-605.
  6. Sanbe, A. et al. (1999) J. Biol. Chem. 274, 21085-21094.

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