Human/Mouse/Rat Vimentin Antibody

Catalog # Availability Size / Price Qty
MAB21052-100
MAB21052-SP
Detection of Human, Mouse, and Rat Vimentin by Western Blot.
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Human/Mouse/Rat Vimentin Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human Vimentin in direct ELISAs. Detects human, mouse, and rat Vimentin in Western blots.
Source
Monoclonal Mouse IgG2A Clone # 979517
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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Immunohistochemistry
5-25 µg/mL
See below
Dual RNAscope ISH-IHC
5-25 µg/mL
See below
Immunocytochemistry
8-25 µg/mL
See below
Knockout Validated
Vimentin is specifically detected in K562 human chronic myelogenous leukemia cell line but is not detectable in Vimentin knockout K562 cell line.
 

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

Western Blot Detection of Human, Mouse, and Rat Vimentin antibody by Western Blot. View Larger

Detection of Human, Mouse, and Rat Vimentin by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MEF mouse embryonic feeder cells, and NR8383 rat alveolar macrophage cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB21052) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 3.

Immunocytochemistry Vimentin antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

Vimentin in HeLa Human Cell Line. Vimentin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB21052) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cytoskeleton. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Vimentin antibody in Human Lymph Node by Immunohistochemistry (IHC-P). View Larger

Vimentin in Human Lymph Node. Vimentin was detected in immersion fixed paraffin-embedded sections of human lymph node using Mouse Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB21052) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Knockout Validated Vimentin Antibody Specificity is Shown by Immunocytochemistry antibody in Knockout Cell Line. View Larger

Vimentin Specificity is Shown by Immunocytochemistry in Knockout Cell Line. Vimentin was detected in immersion fixed K562 human chronic myelogenous leukemia cell line but is not detected in Vimentin knockout (KO) K562 Human Cell Line cell line using Mouse Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB21052) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Dual RNAscope ISH-IHC Compatible Vimentin antibody in Human Tonsil Using Dual RNAscope<sup>®</sup>ISH and IHC. View Larger

Vimentin in Human Tonsil Using Dual RNAscope®ISH and IHC. Vimentin mRNA (red) and protein (green) was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope®Probe (Catalog # 310441) followed by immunohistochemistry using R&D Systems Mouse Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog# MAB21052) at 5ug/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC001). Tissue was stained using ACD RNAscope®2.5 HD Duplex Detection Reagents (Catalog # 322500).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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
epididymis secretory sperm binding protein; FLJ36605; VIM; Vimentin

Product Datasheets

FAQs

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Isotype Controls

Mouse IgG2A Isotype Control

Reconstitution Buffers

Secondary Antibodies

Reviews for Human/Mouse/Rat Vimentin Antibody

Average Rating: 4.8 (Based on 4 Reviews)

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Human/Mouse/Rat Vimentin Antibody
By Anonymous on 11/09/2021
Application: IHC Sample Tested: Ovarian cancer tissue Species: Mouse

Human/Mouse/Rat Vimentin Antibody
By Anonymous on 06/18/2021
Application: IHC Sample Tested: Cancer Tissue Species: Mouse

Human/Mouse/Rat Vimentin Antibody
By Anonymous on 02/20/2021
Application: WB Sample Tested: Pancreatic cancer cells Species: Human

Human/Mouse/Rat Vimentin Antibody
By Anonymous on 07/16/2018
Application: ICC/IF Sample Tested: Pancreatic cancer cells Species: Mouse