Vimentin Antibody (812)

Knockout Validated: Vimentin Antibody (812) [NBP2-43560] - Wild-type (WT) and Vimentin knockout (KO) 293T cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with Vimentin antibody (812) diluted at 1:5000. HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Immunohistochemistry-Frozen: Vimentin Antibody (812) [NBP2-43560] - Frozen sectioned E13.5 Rat brain. Green: Vimentin protein stained by Vimentin antibody [812] diluted at 1:250.Red: beta Tubulin 3/ TUJ1, a mature neuron marker, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:250. Blue: Fluoroshield with DAPI.

Immunohistochemistry-Frozen: Vimentin Antibody (812) [NBP2-43560] - Analysis of Frozen section of embryonic mouse brain (mE18.5). Red: Vimentin antibody [812] diluted at 1:250. Blue: DAPI.

Flow Cytometry: Vimentin Antibody (812) [NBP2-43560] - An intracellular stain was performed on Jurkat cells with NBP2-43560AF00 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

Western Blot: Vimentin Antibody (812) [NBP2-43560] - Whole cell extract (30 ug) was separated by 10% SDS-PAGE, and the membrane was blotted with Vimentin antibody [812] ) diluted at 1:75000.

Immunohistochemistry-Paraffin: Vimentin Antibody (812) [NBP2-43560] - Rat ovary. Vimentin antibody [812] diluted at 1:250. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Western Blot: Vimentin Antibody (812) [NBP2-43560] - Analysis of A. 30 ug 293T whole cell lysate/extract B. 30 ug HeLa whole cell lysate/extract 10 % SDS-PAGE Vimentin antibody [812] dilution: 1:1000

Western Blot: Vimentin Antibody (812) [NBP2-43560] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with Vimentin antibody [812] diluted at 1:3000.

Western Blot: Vimentin Antibody (812) [NBP2-43560] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with Vimentin antibody [812] diluted at 1:5000.

Knockdown Validated: Vimentin Antibody (812) [NBP2-43560] - Non-transfected (-) and transfected (+) 293T whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with Vimentin antibody [GT812].

Sandwich ELISA: Vimentin Antibody (812) [NBP2-43560] - Sandwich ELISA detection of recombinant full-length Vimentin protein using Vimentin Antibody (812) as capture antibody at concentration of 5 ug/mL. HRP-conjugated rabbit IgG antibody was diluted at 1:10000 and used to detect the primary antibody.

• Reactivity: Hu, Mu, Rt, Bv, Ca, Ch, Pm, Gp
• Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, S-ELISA, KD, KO
• Clone: 812
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Vimentin Antibody (812) Summary

• Immunogen: Recombinant protein encompassing a sequence within the center region of human Vimentin. The exact sequence is proprietary.
• Predicted Species: Chicken (92%), Canine (98%), Bovine (98%), Guinea Pig (97%), Chimpanzee (99%). Backed by our 100% Guarantee.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: VIM
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:100-1:1000
  • Immunohistochemistry-Frozen 1:100-1:1000
  • Immunohistochemistry-Paraffin 1:100-1:1000
  • Immunoprecipitation
  • Knockdown Validated
  • Knockout Validated
  • Sandwich ELISA
  • Western Blot 1:500-1:75000
• Application Notes: Flow cytometry/FACS Assay-dependent dilution, ICC/IF Assay-dependent dilution.
• Theoretical MW: 54 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Xenopus laevis (84%).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein G purified

Alternate Names for Vimentin Antibody (812)

• epididymis secretory sperm binding protein
• FLJ36605
• VIM
• Vimentin

Background

Vimentin is a type III intermediate filament (IF) protein, encoded by the VIM gene, that plays an essential role in cytoskeletal function and is largely expressed by mesenchymal cell such as endothelial cells, fibroblasts, and immune cells (1). Human vimentin monomer protein is 466 amino acids (aa) in length with a theoretical molecular weight of ~54 kDa (2). Vimentin protein contains a central rod domain composed of four alpha-helices which is flanked by an N-terminal head domain and C-terminal tail domain (1,3). Vimentin monomers dimerize to form more complex structures resulting in the assembly of eight tetramers, referred to as unit-length filaments, which then assemble into full-length intermediate filaments that are 10-12 nm wide (1,3,4).

Activated macrophages have been shown to secrete phosphorylated vimentin which can be stimulated by a variety of pathophysiological factors including oxidized low-density lipoproteins and TNF-alpha or inhibited by IL-10 (1). The vimentin protein is often expressed at the cell surface playing a role in cell-cell interactions, tissue damage and repair, immune response, and pathogen recognition (1). Vimentin functions in many cytoskeletal processes including cell migration, which is highlighted by its upregulation during epithelial-to-mesenchymal transition (EMT) (4,5). Vimentin is a commonly used marker for EMT and is expressed by many tumor types (5). For example, high metastasis of oral squamous cell carcinomas also showed high vimentin positive expression in immunohistochemical staining analysis (5). A number of vimentin targeting compounds are in cancer-related clinical trials, however, given the multifunctional role of vimentin, the effect of inhibition on non-malignant cells needs to be thoroughly examined (5).

References

1. Ramos, I., Stamatakis, K., Oeste, C. L., & Perez-Sala, D. (2020). Vimentin as a Multifaceted Player and Potential Therapeutic Target in Viral Infections. International Journal of Molecular Sciences. https://doi.org/10.3390/ijms21134675

2. Uniprot (P08670)

3. Morrow, C. S., & Moore, D. L. (2020). Vimentin's side gig: Regulating cellular proteostasis in mammalian systems. Cytoskeleton (Hoboken, N.J.). https://doi.org/10.1002/cm.21645

4. van Bodegraven, E. J., & Etienne-Manneville, S. (2020). Intermediate filaments against actomyosin: the david and goliath of cell migration. Current Opinion in Cell Biology. https://doi.org/10.1016/j.ceb.2020.05.006

5. Strouhalova, K., Prechova, M., Gandalovicova, A., Brabek, J., Gregor, M., & Rosel, D. (2020). Vimentin Intermediate Filaments as Potential Target for Cancer Treatment. Cancers. https://doi.org/10.3390/cancers12010184

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Vimentin Antibody (NBP2-43560)(5)

Publications using NBP2-43560

• Gabra N Circumferential tracheal replacement using 3D polycaprolactone scaffold and adipose derived mesenchymal stromal cells Thesis Jan 1 2021 (IHC-Fr)
• Poole JA, Gaurav R, Schwab A et al. Post-endotoxin exposure-induced lung inflammation and resolution consequences beneficially impacted by lung-delivered IL-10 therapy Scientific reports Oct 15 2022 [PMID: 36243830] (Flow, Mouse)
• Manzer HS, Villarreal RI, Doran KS Targeting the BspC-vimentin interaction to develop anti-virulence therapies during Group B streptococcal meningitis PLoS pathogens Mar 1 2022 [PMID: 35316308]
• Kaul A, Kuthethur R, Ishida Y et al. Molecular Insights into the Antistress Potentials of Brazilian Green Propolis Extract and Its Constituent Artepillin C Molecules Dec 23 2021 [PMID: 35011307]
• Swoger M, Gupta S, Charrier EE et al. Vimentin Intermediate Filaments Mediate Cell Morphology on Viscoelastic Substrates ACS applied bio materials Jan 7 2022 [PMID: 34995457] (ICC/IF)

Bioinformatics

• Gene Symbol: VIM