Human/Mouse/Rat Vimentin Antibody MAB2105: R&D Systems

Human/Mouse/Rat Vimentin Antibody

(3 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Vimentin antibodies are ideal for immunocytochemistry colocalization studies in intermediate filaments. Detects human, mouse and rat 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 Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1-2 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • 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
Detection of Human Vimentin by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Vimentin at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Mouse and Rat Vimentin by Western Blot. Western blot shows lysates of MEF mouse embryonic feeder cells, NIH‑3T3 mouse embryonic fibroblast cell line, Rat‑2 rat embryonic fibroblast cell line, and NR8383 rat alveolar macrophage cell line. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Vimentin at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Vimentin in NTera‑2 Human Cell Line. Vimentin was detected in immersion fixed NTera‑2 human testicular embryonic carcinoma cell line using Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (yellow; Catalog # NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
Vimentin in A549 Human Cell Line. Vimentin was detected in immersion fixed A549 human lung carcinoma cell line using Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Rat IgG Secondary Antibody (green; Catalog # NL015) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
Vimentin in Mouse Cortical Stem Cells. Vimentin was detected in immersion fixed mouse cortical stem cells using Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoskeleton. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
Vimentin in Rat Cortical Stem Cells. Vimentin was detected in immersion fixed rat cortical stem cells using Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoskeleton. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Vimentin by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Vimentin at approximately 58 kDa (as indicated) using 10 µg/mL of Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody
(Catalog # MAB2105) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
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. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Combined CSL and p53 downregulation promotes cancer-associated fibroblast activation.
    Authors: Procopio M, Laszlo C, Al Labban D, Kim D, Bordignon P, Jo S, Goruppi S, Menietti E, Ostano P, Ala U, Provero P, Hoetzenecker W, Neel V, Kilarski W, Swartz M, Brisken C, Lefort K, Dotto G
    Nat Cell Biol, 2015;17(9):1193-204.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Not specified
  2. GLI1, CTNNB1 and NOTCH1 protein expression in a thymic epithelial malignancy tissue microarray.
    Authors: Riess J, West R, Dean M, Klimowicz A, Neal J, Hoang C, Wakelee H
    , 2015;0(0):.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  3. Development of a reconstructed cornea from collagen-chondroitin sulfate foams and human cell cultures.
    Authors: Vrana NE, Builles N, Justin V, Bednarz J, Pellegrini G, Ferrari B, Damour O, Hulmes DJ, Hasirci V
    Invest. Ophthalmol. Vis. Sci., 2008;49(12):5325-31.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
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