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Vimentin Antibody (2D1)

Images

 
Genetic Strategies: 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. ...read more
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 ...read more
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, ...read more
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 ...read more
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 ...read more
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 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 ...read more
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 ...read more

Product Details

Summary
Reactivity Hu, Rt, Mu(-)Species Glossary
Applications WB, Simple Western, ICC/IF, IHC, KO
Clone
2D1
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1 mg/ml
Validated by:
     

Genetic Strategies

   

Order Details

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Vimentin Antibody (2D1) Summary

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
Localization
Cytoplasm
Marker
Mesenchymal Cells Marker
Isotype
IgG2a
Clonality
Monoclonal
Host
Mouse
Gene
VIM
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry/ Immunofluorescence 1:1000
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Paraffin
  • Knockout Validated
  • 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.
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.
Reviewed Applications
Read 1 Review rated 5
using
NBP1-92687 in the following applications:

Publications
Read Publications using
NBP1-92687 in the following applications:

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.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
50% PBS, 50% glycerol
Preservative
0.035% Sodium Azide
Concentration
1 mg/ml
Purity
Protein G purified

Alternate Names for Vimentin Antibody (2D1)

  • 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.

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Publications for Vimentin Antibody (NBP1-92687)(11)

We have publications tested in 3 confirmed species: Human, Rat,  Human.

We have publications tested in 4 applications: ICC/IF, IF/IHC, IHC-P, WB.


Filter By Application
ICC/IF
(4)
IF/IHC
(1)
IHC-P
(2)
WB
(2)
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Human
(7)
Rat
(1)
 Human
(1)
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Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using NBP1-92687 Applications Species
Duan X, Weng H, Shen G et al. Preliminary study on the expression of UHRF1 in early oral squamous cell carcinoma and its clinical prognostic value Research Square 2023-02-03 (IHC-P, Human) IHC-P Human
Chou CW, Hung CN, Chiu CH et al. Phagocytosis-initiated tumor hybrid cells acquire a c-Myc-mediated quasi-polarization state for immunoevasion and distant dissemination Nature communications 2023-10-17 [PMID: 37848444]
Manhas A, Tripathi D, Jagavelu K Involvement of HIF1?/Reg protein in the regulation of HMGB3 in myocardial infarction Vascular pharmacology 2023-07-17 [PMID: 37467910] (ICC/IF, Rat) ICC/IF Rat
Monleón-Guinot I, Milian L, Martínez-Vallejo P et al. Morphological Characterization of Human Lung Cancer Organoids Cultured in Type I Collagen Hydrogels: A Histological Approach International journal of molecular sciences 2023-06-14 [PMID: 37373279] (ICC/IF, Human)

Details:
1:500 dilution
ICC/IF Human
Wang W, Lin Y, Zhang F et al. Single cell transcriptomic analysis of human amnion identifies cell-specific signatures associated with membrane rupture and parturition Cell Biosci. 2022-05-18 [PMID: 35585644] (ICC/IF, Human) ICC/IF  Human
Hamdan F, YlOsmAki E, Chiaro J et al. Novel oncolytic adenovirus expressing enhanced cross-hybrid IgGA Fc PD-L1 inhibitor activates multiple immune effector populations leading to enhanced tumor killing in vitro, in vivo and with patient-derived tumor organoids J Immunother Cancer 2021-11-01 [PMID: 34362830] (ICC/IF, Human) ICC/IF Human
Chandramohan Y, Jeganathan K, Sivanesan S et al. Assessment of human ovarian follicular fluid derived mesenchymal stem cells in chitosan/PCL/Zn scaffold for bone tissue regeneration Life Sci 2020-10-06 [PMID: 33031825] (WB, Human) WB Human
Park S, Song CS, Lin CL et al. Inhibitory Interplay of SULT2B1b Sulfotransferase with AKR1C3 Aldo-keto Reductase in Prostate Cancer Endocrinology 2020-01-02 [PMID: 31894239]
Chen, YC;Ou, MC;Fang, CW;Lee, TH;Tzeng, SL; High Glucose Concentrations Negatively Regulate the IGF1R/Src/ERK Axis through the MicroRNA-9 in Colorectal Cancer Cells 2019-04-08 [PMID: 30965609] (WB, Human) WB Human
Scimeca M, Urbano N, Bonfiglio R et al. Breast osteoblast-like cells: a new biomarker for the management of breast cancer. Br. J. Cancer. 2018-10-17 [PMID: 30327566] (IHC-P, Human) IHC-P Human
Show All 11 Publications.

Review for Vimentin Antibody (NBP1-92687) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Mouse.

Reviews using NBP1-92687:
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Immunohistochemistry-microslice
(1)
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Mouse
(1)
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5
reviewed by:
Verified Customer
Immunohistochemistry-microslice Mouse 07/27/2017
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Summary

ApplicationOther
Sample TestedMouse cortex
SpeciesMouse
Lot041813

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Vimentin Antibody (NBP1-92687). (Showing 1 - 2 of 2 FAQs).

  1. 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?
    • 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.
  2. Hi, what exactly does Mu(-) refer to?
    • (-) refers to a negative species reactivity. In this case, the antibody reacts to human and rat Vimentin but not to mouse Vimentin

Secondary Antibodies

 

Isotype Controls

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Research Areas for Vimentin Antibody (NBP1-92687)

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Blogs on Vimentin. Showing 1-10 of 14 blog posts - Show all blog posts.

Taking Biomarker Discovery From 2D to 3D: Increased Biological Activity of EVs Isolated From 3D Prostate Cancer Cultures
Jamshed Arslan, Pharm D, PhD Tissues within the human body are made of a three-dimensional (3D) arrangement of cells working together to perform vital functions. The commonly used 2D monolayer cultures have limited ...  Read full blog post.


  Read full blog post.

Antibody treatment can generate microglia-like cells from bone marrow
By Jennifer Sokolowski, MD, PhD.Microglia play important roles in the brain in both homeostatic and pathological conditions, acting to clear debris and dying cells. There is evidence to suggest that microglial dys...  Read full blog post.

Stemness is responsible for onset and metastasis of colorectal cancer
By Jamshed Arslan, Pharm. D., PhD. Colorectal cancer stem cells are a rare subpopulation of colorectal cancer cells that can self-renew and initiate and sustain tumor growth when transplanted into an animal host.1,2 C...  Read full blog post.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

The use of actin as a loading control in research on fruiting-body development and vegetative growth in Sordaria macrospora research
Sordaria macrospora is a filamentous fungus that serves as very useful system for scientific research due to a short life cycle and easy manipulation.  Just like any other model organism, it is important to have an effective loading control to va...  Read full blog post.

Alpha-smooth muscle actin and the modulation of endothelial and epithelial cell biology
Actin is essential for a wide range of cell functions, ranging from cell division and chromatin remodeling to vesicle trafficking and maintenance of cellular structure. In fact, mislocalization of actin to cell junctions during development leads t...  Read full blog post.

Epithelial-Mesenchymal Transition (EMT) Markers
Epithelial-Mesenchymal Transition (EMT) is the trans-differentiation of stationary epithelial cells into motile mesenchymal cells. During EMT, epithelial cells lose their junctions and apical-basal polarity, reorganize their cytoskeleton, undergo a...  Read full blog post.

CD63: is it pro-metastatic or anti-metastatic?
CD63 is a type II membrane protein belonging to tetraspanin superfamily and it play key roles in the activation of several cellular signaling cascades along with acting as TIMP1 receptor. It is expressed by activated platelets, monocytes,...  Read full blog post.

Vimentin: Regulating EMT and Cancer
Vimentin, a member of the intermediate filament (IF) family, is a protein responsible for maintaining cellular integrity and reducing damage caused by stress. The vimentin protein is ubiquitously expressed in normal mesenchymal cells, and recent resea...  Read full blog post.

Showing 1-10 of 14 blog posts - Show all blog posts.
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Verified Customer
07/27/2017
Application: Immunohistochemistry-microslice
Species: Mouse

Bioinformatics

Gene Symbol VIM
Entrez
Uniprot