Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Rat, Mouse (Negative)

Cited:

Human, Rat

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Knockout Validated, Simple Western, Western Blot

Cited:

IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 2D1

Concentration

1 mg/ml

View all Vimentin Antibodies »

Product Specifications

Immunogen

Full length recombinant human Vimentin Antibody expressed in and purified from E. coli. [UniProt# P08670]

Epitope

amino acids 409-425 of human Vimentin (UniProt Accession P08670): SRISLPLPNFSSLNREL

Reactivity Notes

Clones 2D1 (NBP1-92687) and 2A52 (NBP1-92688) both failed to detect the target in mouse tissues although they work well on human and rat samples. This allowed us to firmly map the epitope for both antibodies to the peptide SRISLPLPNFSSLNREL, amino acids 409-425 of the human sequence. This peptide is located at the beginning of the non-helical "tail" region of the molecule and the peptide is totally conserved between human and rat and in most mammalian species, including cow, pig, horse, camel, and many monkeys. Interestingly mouse has the peptide SRISLPLPTFSSLNREL divergent by one amino acid, and neither clones bind this peptide. As a result these antibodies can be used to identify human or rat cells in mouse cultures or tissues and may work with other species that also contain this peptide.

Localization

Cytoplasm

Marker

Mesenchymal Cells Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Theoretical MW

53.6 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.

Scientific Data Images for Vimentin Antibody (2D1)

Knockout Validated: Vimentin Antibody (2D1) [NBP1-92687]

Knockout Validated: Vimentin Antibody (2D1) [NBP1-92687]

Knockout Validated: Vimentin Antibody (2D1) [NBP1-92687] - Western blot shows lysates of K562 human Chronic Myelogenous Leukemia parental cell line and Vimentin knockout (KO) K562 cell line. PVDF membrane was probed with 1:10,000 of Mouse Anti-Human Vimentin Monoclonal Antibody (Catalog # NBP1-92687) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog #HAF018). Specific band was detected for Vimentin at approximately 55 kDa (as indicated) in the parental K562 cell line, but is not detectable in the knockout K562 cell line. This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: Vimentin Antibody (2D1) [NBP1-92687]

Immunocytochemistry/ Immunofluorescence: Vimentin Antibody (2D1) [NBP1-92687]

Immunocytochemistry/Immunofluorescence: Vimentin Antibody (2D1) [NBP1-92687] - View of mixed neuron/glial cultures stained with NBP1-92687 (green) and the GFAP rabbit polyclonal (NB300-141, red). Vimentin is expressed alone in fibroblastic and endothelial cells, which are the flattened cells in the middle of the image which appear green. Astrocytes may express primarily GFAP, or GFAP and Vimentin, and so appear red (GFAP only) or golden yellow (GFAP and Vimentin). In cells which express both GFAP and Vimentin, the two proteins assemble to produce heteropolymer filaments.
Western Blot: Vimentin Antibody (2D1) [NBP1-92687]

Western Blot: Vimentin Antibody (2D1) [NBP1-92687]

Western Blot: Vimentin Antibody (2D1) [NBP1-92687] - Analysis of different cell lysates using mouse mAb to vimentin, NBP1-92687, dilution 1:10,000 in green: [1] protein standard (red), [2] HEK293, [3] HeLa, [4] SH-SY5Y, [5] COS-1, and [6] C6 cells. The band at about 50kDa mark corresponds to the vimentin protein.
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Applications for Vimentin Antibody (2D1)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000

Immunohistochemistry

1:1000

Simple Western

1:100

Western Blot

1:10000
Application Notes
This Vimentin (2D1) antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western blot, where a band can be seen at approximately 50 kDa. Use in IHC-P was reported in scientific literature (PMID: 30327566).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NBP1-92687 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50% PBS, 50% glycerol

Preservative

0.035% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Vimentin

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

Alternate Names

VIM

Entrez Gene IDs

7431 (Human)

Gene Symbol

VIM

UniProt

P08670

Additional Vimentin Products

Product Documents for Vimentin Antibody (2D1)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Vimentin Antibody (2D1)

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.

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