Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Rat, Mouse (Negative)

Cited:

Human, Rat

Applications

Validated:

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

Cited:

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

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 2D1

Format

BSA Free
View all Vimentin Primary 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.

Description

Novus Biologicals Knockout (KO) Validated Mouse Vimentin Antibody (2D1) - BSA Free (NBP1-92687) is a monoclonal antibody validated for use in IHC, WB, ICC/IF and Simple Western. Anti-Vimentin Antibody: Cited in 10 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for Vimentin Antibody (2D1) - BSA Free

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

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

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

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

Immunocytochemistry/Immunofluorescence: Vimentin Antibody (2D1) [NBP1-92687] - Analysis of cortical neuron-glial cell cultures from E20 rat stained with mouse mAb to vimentin, NBP1-92687, dilution 1:2,000 in red, and costained with chicken pAb to glial fibrillary acidic protein (GFAP), dilution 1:5,000, in green. The blue is DAPI staining of nuclear DNA. Fibroblastic and other developing cells express only vimentin and appear red. Astrocytes that express GFAP only are green while those that express both GFAP and vimentin appear golden yellow.
Immunohistochemistry-Paraffin: Vimentin Antibody (2D1) [NBP1-92687]

Immunohistochemistry-Paraffin: Vimentin Antibody (2D1) [NBP1-92687]

Immunohistochemistry-Paraffin: Vimentin Antibody (2D1) [NBP1-92687] - Human breast cancer tissue stained for Vimentin (white) and countersatined with DAPI (blue). Alexa Fluor 750 version of antibody used (NBP1-92687AF750). Image from verified customer review.
Immunohistochemistry: Vimentin Antibody (2D1) [NBP1-92687]

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

Immunohistochemistry: Vimentin Antibody (2D1) [NBP1-92687] - Mouse cortex microslice section (from postnatal day 3) was immunostained with (1:500 dil.). This image was submitted via customer Review.
Simple Western: Vimentin Antibody (2D1) [NBP1-92687]

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

Simple Western: Vimentin Antibody (2D1) [NBP1-92687] - Simple Western lane view shows a specific band for Vimentin in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Vimentin Antibody (2D1)

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

Western Blot: Vimentin Antibody (2D1) [NBP1-92687] - High glucose (HG) concentrations induced epithelial-to-mesenchymal transition protein expression & enhanced migration activity in colorectal cancer (CRC) cells. SW480 (low metastatic potential) & SW620 (high metastatic potential) cells were cultured in different concentrations of glucose (normal: NG; HG; & osmotic control: NG + l-glucose). (A) Morphological change occurred from epithelial to mesenchymal type in the HG-concentration group. (B) HG concentration caused downregulation of E-cadherin & upregulation of N-cadherin, beta CTN, & vimentin, but c-myc was unchanged, as detected using Western blotting. beta -actin was evaluated as an internal control. (C,D) Wound healing assay showed that HG concentration promoted cell motility in SW480 & SW620 CRC cells after 48 & 72 h of culture, compared with the NG & NG + l-glucose groups. (E) In a Transwell migration assay, 3.5 × 105 SW480 & SW620 CRC cells were plated onto a 24-well plate & cultured in NG & HG-concentration medium for 96 h. HG concentration promoted cell motility in SW480 & SW620 cells. NG + l-glucose cells were evaluated as ostomic controls. (F) These data show that HG concentration caused upregulation of p-IGF1R in CRC. In addition, HG concentration promoted IGF1R downstream signaling, including p-Src & p-ERK; these proteins were increased when CRC cells were cultured in HG-concentration medium. Levels of beta -actin were evaluated as loading controls. Statistically significant differences between the two groups were judged using Student’s t-tests; * p < 0.05, ** p < 0.005, *** p < 0.001; n.s. = nonsignificant. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30965609), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Vimentin Antibody (2D1) - BSA Free

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.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:100, apparent MW was 59 kDa. 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

Format

BSA Free

Preservative

5mM 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) - BSA Free

Certificate of Analysis

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Product Specific Notices for Vimentin Antibody (2D1) - BSA Free

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.

Citations for Vimentin Antibody (2D1) - BSA Free

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Customer Images

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  • Vimentin Antibody (2D1)
    Name: Anonymous
    Application: Immunohistochemistry-microslice
    Sample Tested: Mouse cortex
    Species: Mouse
    Verified Customer | Posted 07/27/2017
    Mouse cortex microslice section (from postnatal day 3) was immunostained with Vimentin Antibody (2D1) (1:500 dil.)
    Vimentin Antibody (2D1) - BSA Free NBP1-92687

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FAQs for Vimentin Antibody (2D1) - BSA Free

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    • A: (-) refers to a negative species reactivity. In this case, the antibody reacts to human and rat Vimentin but not to mouse Vimentin

    • Q: I am planning on using mouse monoclonal Vimentin Antibody (2D1), NBP1-92687 on mouse fibroblasts. Has the antibody been tested for cross-reactivity with endogenous IgG? Do I have to expect strong background?
      A: If the proposed samples are mouse fibroblasts grown in tissue culture, there should be no problem as any endogenous mouse IgG would be diluted out after a few days in culture, as the cells are fed. Western blotting or IHC of pancreatic tissue would be more of a problem, as this would contain some endogenous IgG, though much of this can be removed by perfusing the animal with PBS prior to taking out the pancreas. The endogenous mouse IgG is mostly in blood vessels and may show up in IHC, though various kits are available which are supposed to address this issue. On western blots, the mouse IgG usually shows up as a somewhat diffuse band at about 55 kDa (the IgG heavy chains; most anti-mouse reagents bind primarily the heavy chain) and in some cases a faint band at about 20 kDa (the IgG light chains). Vimentin should appear as a sharp and clean single band at 50 kDa, so should be easily distinguished from any endogenous mouse IgG. This issue could be avoided entirely by using an alternative anti-vimentin antibody with a different host.
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