Overview

  • Product nameAnti-Vimentin antibody [RV202]
    See all Vimentin primary antibodies
  • Description
    Mouse monoclonal [RV202] to Vimentin
  • SpecificityThis antibody reacts exclusively with vimentin
  • Tested applicationsSuitable for: Flow Cyt, ICC, IHC-Fr, WB, IHC-FoFr, IP, IHC-P, ICC/IF, IHC (PFA fixed)more details
  • Species reactivity
    Reacts with: Mouse, Rat, Goat, Chicken, Hamster, Cow, Dog, Human, Xenopus laevis, Monkey, Zebrafish
  • Immunogen

    Fusion protein corresponding to Human Vimentin. RV202 is a mouse monoclonal IgG1 antibody derived by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells from a BALB/c mouse immunized with a vimentin extract of bovine lens.

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab8978 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC 1/20.
IHC-Fr Use at an assay dependent concentration. Recommended range is 1/100 - 1/200 for Immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection. For PFA fixed tissue use at 1/1000.
WB 1/100 - 1/1000. Detects a band of approximately 57 kDa.
IHC-FoFr 1/500.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 20097175
IHC (PFA fixed) 1/1000.

Target

  • FunctionVimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
    Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
  • Tissue specificityHighly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.
  • Involvement in diseaseCataract 30
  • Sequence similaritiesBelongs to the intermediate filament family.
  • DomainThe central alpha-helical coiled-coil rod region mediates elementary homodimerization.
    The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
  • Post-translational
    modifications
    Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33.
    O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.
    S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
  • Cellular localizationCytoplasm.
  • Information by UniProt
  • Database links
  • FormVimentin is found in connective tissue and in the cytoskeleton.
  • Alternative names
    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Anti-Vimentin antibody [RV202] images

  • ab8978 were fixed with paraformaldehyde, permeabilized with PBS and 0.5% Triton ×100 and blocking with 0.1% BSA + 10% Goat Serum at 250C for 30 minutes was performed. Samples were incubated with primary antibody (1/250: in PBS, 0.1% BSA and 10% Goat Serum) for 12 hours at 4°C. An Alexa Fluor®594-conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

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  • ab8978 staining Vimentin in Human fetal kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with CAS-Block for 1 hour at 25°C; antigen retrieval was by heat mediation using OmniPrep (pH 9). Samples were incubated with primary antibody (1/500) for 1 hour at 25°C. An Alexa Fluor® 555-conjugated Donkey polyclonal (1/200) was used as the secondary antibody.

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  • Immunofluorescence staining images of 9 day old zebrafish embryos.
    ab8978 reacts with in connective tissue cells and bloodvessels. Frozen sample treated with Acetone:Methanol 1:1, antibody diluted 1/100 and incubated for 45 minutes at room temperature.

  • Overlay histogram showing HeLa cells stained with ab8978 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8978, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This anti-Vimentin antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

  •   ab8978 staining Vimentin - Neural Stem Cell Marker in Human Colon fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol, permeabilized in 0.1% Triton and blocked with 0.25% serum free protein blocker for 20 minutes at 28°C. Samples were incubated with primary antibody (1/100 in antibody diluent) for 2 hours at 28°C. ab6785 Goat polyclonal anti-Mouse IgG - H&L (FITC) (1/800) was used as the secondary antibody. Nuclei were counterstained with propidium iodide.

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  • ab8978 staining Vimentin in Dog soft tissue sarcoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 15% serum for 1 hour at 20°C; antigen retrieval was by heat mediation in a Tris/EDTA pH9 buffer. Samples were incubated with primary antibody (1/100 in TBS) for 18 hours at 20°C. A Alexa Fluor® 647-conjugated Goat anti-mouse IgG polyclonal (1/400) was used as the secondary antibody.

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  • IHC-Fr image of Ed18 rat stained with ab8978. Fresh frozen sections  were  incubated in  10% normal donkey serum in 0.1% PBS- and 0.3% triton X100  for 1h to permeabilise the tissues and block non-specific protein-protein interactions. The sectons were then incubated with the ab8978 (1µg/ml) and ferroportin overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 568 (red) donkey anti-mouse at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. Vimentin expressed in the gut muscles.

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  • IHC-FoFr image of vimentin staining on rat injured cortical sections using ab8978 (1:500). The brain was perfusion fixed using 4% PFA and the sections were permeabilized using 0.1% TritonX in 0.1% PBS. The sections were then blocked using 10% donkey serum for 1 hour at 24°C. ab8978 was diluted 1:500 and incubated with sections for 24 hours using 4°C. The secondary antibody used was donkey polyclonal to rabbit IgG conjugated to Alexa Fluor 488.

  • ab8978 vimentin staining of a tonsilar lymphoma. Note that the epithelium (at the left) is negative.

  •  ab8978 staining Vimentin in bovine chromospheres by ICC/IF (immunocytochemistry/immunofluorescence). Cells were PFA fixed and permeabilized in 0.3% Triton X-100. The primary antibody (1/500) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 568-conjugated goat anti-mouse IgG polyclonal (1/500) ab175473 was used as the secondary.  

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  • ab8978 vimentin immunofluorescent staining of cultured bovine lens epithelial cells

  •  ab8978 staining vimentin in human pancreatic adenocarcinoma cells by immunocytochemistry/ immunofluorescence. Cells were PFA fixed and permeabilized in 0.2% Triton X prior to blocking in 3% BSA for 30 minutes at 24°C. The primary antibody was diluted 1/200 and incubated with the sample for 16 hours at 21°C. Alexa fluor® 488 mouse polyclonal to mouse Ig, diluted 1/300 was used as the secondary antibody.

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  • ab8978staining Vimentin in human lung tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and blocking with 5% commercially available blocking agent was performed at 370C  for 15 minutes. The sample was incubated with primary antibody (1/250) at 370C for 1 hour. A HRP-conjugated Goat polyclonal to mouse IgG was used as secondary antibody at 1/1000 dilution.

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References for Anti-Vimentin antibody [RV202] (ab8978)

This product has been referenced in:
  • Li C  et al. High-Content Functional Screening of AEG-1 and AKR1C2 for the Promotion of Metastasis in Liver Cancer. J Biomol Screen 21:101-7 (2016). Read more (PubMed: 26318406) »
  • Fay ME  et al. Cellular softening mediates leukocyte demargination and trafficking, thereby increasing clinical blood counts. Proc Natl Acad Sci U S A 113:1987-92 (2016). Read more (PubMed: 26858400) »

See all 42 Publications for this product

Product Wall

The exact position of epitope sequence recognized by anti vimentin antibody (clone RV202) is not determined however, the antibody must recognize an evolutionary highly conserved epitope, since it shows a broad spectrum of species cross reactivity (from...

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Indeed, a mouse-on mouse staining can be quite tricky; however, it is not impossible. The background will depend on the fixation method ( perfusion fixed tissue vs. normally fixed tissue), on the tissue itself ( ...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (fibroblast)
Permeabilization No
Specification fibroblast
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Aug 19 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (melanoma cell line xenograft)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification melanoma cell line xenograft
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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Submitted Dec 21 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (canine soft tissue sarcoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification canine soft tissue sarcoma
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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Submitted Dec 18 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (fetal kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: commercial - ZYTOMED's OmniPrep (pH 9)
Permeabilization Yes - 0.05% tween-20
Specification fetal kidney
Blocking step commercial - Invitrogen's CAS-Block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Dec 11 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Sample Human Cell (iPS-derived cardiofibroblast)
Specification iPS-derived cardiofibroblast
Permeabilization Yes - 0.2% triton X
Fixative Formaldehyde
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Submitted Aug 04 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (gel 10%)
Sample Human Cell lysate - whole cell (breast cancer cell line MDAMB231)
Specification breast cancer cell line MDAMB231
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
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Submitted Dec 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Sample Rat Cell (Rat Brain post injury)
Specification Rat Brain post injury
Permeabilization Yes - 0.1% TritonX in 0.1% PBS
Fixative Paraformaldehyde
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Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Sep 27 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (stromal cell)
Specification stromal cell
Fixative Formaldehyde
Permeabilization Yes - Saponin
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Apr 02 2013

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