Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-Vimentin (Ser56) Antibody #3877

Applications Reactivity MW (kDa) Source
W IF-IC H M R Mk 57 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Vimentin (Ser56) Antibody detects endogenous levels of vimentin only when phosphorylated at Ser56.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser56 of human vimentin. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types, hydroxyurea-treated (4 mM) (G1/S) or paclitaxel-treated (100 nM) (G2/M) for 20 hours, using Phospho-Vimentin (Ser56) Antibody (upper). β-Actin Antibody #4967 was used as a loading control (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Phospho-Vimentin (Ser56) Antibody (green) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Background

The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments and microtubules. Major types of intermediate filaments are distinguished and expressed in particular cell types: cytokeratins (epithelial cells), glial fibrillary acidic protein, GFAP (glial cells), desmin (skeletal, visceral and certain vascular smooth muscle cells), vimentin (mesenchyme origin) and neurofilaments (neurons). GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). Vimentin is present in sarcomas, but not carcinomas, and its expression is examined in conjunction with that of other markers to distinguish between the two (3). Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli helps to coordinate various signaling pathways (4). Phosphorylation of vimentin at Ser56 in smooth muscle cells regulates structural arrangement of vimentin filaments in response to serotonin (5,6). Remodeling of vimentin and other intermediate filaments is important during lymphocyte adhesion and migration though the endothelium (7).

During mitosis, CDK1 phosphorylates vimentin at Ser56. This phosphorylation provides a PLK binding site for vimentin-PLK interaction. PLK further phosphorylates vimentin at Ser82 , which might serve as memory phosphorylation site and play a regulatory role in vimentin filament disassembly (8,9).

  1. Eng, L.F. et al. (2000) Neurochem. Res. 25, 1439-1451.
  2. Goebel, H.H. et al. (1987) Acta Histochem. Suppl. 34, 81-93.
  3. Leader, M. et al. (1987) Histopathology 11, 63-72.
  4. Helfand, B.T. et al. (2004) J. Cell Sci. 117, 133-141.
  5. Tang, D.D. et al. (2005) Biochem. J. 388, 773-783.
  6. Fomina, I.G. et al. (1990) Klin. Med. (Mosk.) 68, 125-127.
  7. Nieminen, M. et al. (2006) Nat. Cell Biol. 8, 156-162.
  8. Yamaguchi, T. et al. (2005) J. Cell Biol. 171, 431-436.
  9. Oguri, T. et al. (2006) Genes Cells 11, 531-540.

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