Phospho-Vimentin (Ser56) Antib

3877:

Immunofluorescence , Western Blotting

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

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser56 of human vimentin. Antibodies are purified by peptide affinity chromatography.

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

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^® #4084 (fluorescent DNA dye).

The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are distinguished by their cell specific expression: 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 help to coordinate various signaling pathways (4). Phosphorylation of vimentin at Ser56 in smooth muscle cells regulates the 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 through 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.

• Slawson, C. et al. (2008) Mol Biol Cell 19, 4130-40. Applications: Western Blotting

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• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7075 Anti-biotin, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 3878 Phospho-Vimentin (Ser82) Antibody
• 4967 β-Actin Antibody

For Research Use Only. Not For Use In Diagnostic Procedures.