Human/Mouse/Rat Vimentin Biotinylated Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Vimentin antibodies are ideal for immunocytochemistry colocalization studies in intermediate filaments. The unconjugated antibody detects human Vimentin in Western blots.
  • Source
    Monoclonal Rat IgG2A Clone # 280618
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human Vimentin
    Ser2-Glu466
    Accession # P08670
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
  • Label
    Biotin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
Vimentin in HeLa Human Cell Line. Vimentin was detected in formaldehyde fixed HeLa human cervical epithelial carcinoma cell line using Rat Anti-Human/Mouse/Rat Vimentin Biotinylated Monoclonal Antibody (Catalog # BAM2105) at 8 µg/mL overnight at 4 ° C. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (orange; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to intermediate filaments. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Vimentin

Vimentin is a 57 kDa class III intermediate filament (IF) protein that belongs to the intermediate filament family. It is the predominant IF in cells of mesenchymal origin such as vascular endothelium and blood cells (1-3). The human Vimentin cDNA encodes a 466 amino acid (aa) protein that contains head and tail regions with multiple regulatory Ser/Thr phosphorylation sites, and a central rod domain with three coiled-coil regions separated by linkers (1, 2). Human Vimentin shares 97-98% aa identity with mouse, rat, ovine, bovine, and canine Vimentin. Sixteen Vimentin coiled-coil dimers self-assemble to form intermediate (10-12 nm wide) filaments (4). These filaments then anneal longitudinally to form non-polarized fibers that support cell structure and withstand stress (4). IF fibers are highly dynamic, and half-life depends on the balance between kinase and phosphatase activity. For example, phosphorylation followed by dephosphorylation drives IF disintegration, followed by reorganization during mitosis (1, 5, 6). Interactions of head and tail domains link IFs with other structures such as actin and microtubule cytoskeletons (7). Vimentin is involved in positioning autophagosomes, lysosomes and the Golgi complex within the cell (8). It facilitates cell migration and motility by recycling internalized trailing edge integrins back to the cell surface at the leading edge (9-11). Vimentin helps maintain the lipid composition of cellular membranes, and caspase cleavage of Vimentin is a key event in apoptosis (8, 12). Phosphorylation promotes secretion of Vimentin by TNF-alpha -stimulated macrophages (13). Extracellular Vimentin has been shown to associate with several microbes, and appears to promote an antimicrobial oxidative burst (13, 14). Cell-associated Vimentin can also interact with NKp46 to recruit NK cells to tuberculosis-infected monocytes (15).

  • References:
    1. Omary, M.B. et al. (2006) Trends Biochem. Sci. 31:383.
    2. Ivaska, J. et al. (2007) Exp. Cell Res. 313:2050.
    3. Ferrari, S. et al. (1986) Mol. Cell. Biol. 6:3614.
    4. Sokolova, A.V. et al. (2006) Proc. Natl. Acad. Sci. USA 103:16206.
    5. Eriksson, J.E. et al. (2004) J. Cell Sci. 117:919.
    6. Li, Q-F. et al. (2006) J. Biol. Chem. 281:34716.
    7. Esue, O. et al. (2006) J. Biol. Chem. 281:30393.
    8. Styers, M.L. et al. (2005) Traffic 6:359.
    9. McInroy, L. and A. Maata (2007) Biochem. Biophys. Res. Commun. 360:109.
    10. Nieminen, M. et al. (2006) Nat. Cell Biol. 8:156.
    11. Ivaska, J. et al. (2005) EMBO J. 24:3834.
    12. Byun, Y. et al. (2001) Cell Death Differ. 8:443.
    13. Mor-Vaknin, N. et al. (2003) Nat. Cell Biol. 5:59.
    14. Zou, Y. et al. (2006) Biochem. Biophys. Res. Commun. 351:625.
    15. Garg, A. et al. (2006) J. Immunol. 177:6192.
  • Entrez Gene IDs:
    7431 (Human); 22352 (Mouse); 81818 (Rat)
  • Alternate Names:
    FLJ36605; VIM; Vimentin
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