Overview

  • Product nameAnti-Vimentin antibody [VI-10]
    See all Vimentin primary antibodies
  • Description
    Mouse monoclonal [VI-10] to Vimentin
  • SpecificityThe antibody VI-10 reacts with vimentin, a 57 kDa intermediate filament expressed in variety of mesenchymal and mesodermal cell types.
  • Tested applicationsSuitable for: ICC/IF, IP, WB, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Dog, Human, Pig
  • Immunogen

    Full length native protein (purified) corresponding to Vimentin.

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab20346 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
ICC/IF Use at an assay dependent concentration. Used at a dilution of 1/1000, 1 hr incubation on rat astrocytes (see Abreview).
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
ICC Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.

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 [VI-10] images

  • Human normal skin. Staining is localised to cytoplasm. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Immunocytochemistry/ Immunofluorescence analysis of chicken postnatal erythrocytes labelling Vimentin (green) with ab20346. Tubulin was counterstained red with anti-alpha-tubulin and DAPI was used as a nuclear counterstain. 
  • Immunofluorescence staining of RBL rat basophiilioc cell line with anti-Vimentin (VI--10). Nuclei are stained with DAPI (blue).

     

  •  ab20346 at a 1/1000 dilution staining rat astrocyte cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a goat anti-mouse Alexa-Fluor ® 568 (ab175473).

    This image is courtesy of an Abreview submitted by Randal Moldrich on 8 February 2006

    See Abreview

  • ab20346 at a 1/500 dilution staining mouse dissociated neural precursor cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a goat anti-mouse Alexa-Fluor 568 (red stain) ab175473>ab175473). The image also shows a blue nuclear counterstain. Confocal image is shown with 2x zoom detail in top right corner.

    This image is courtesy of an Abreview submitted by Randal Moldich on 22 February 2006.

    See Abreview



  • Predicted band size : 54 kDa

    Image from Gao MQ et al, J Cell Sci. 2010 Oct 15;123(Pt 20):3507-14. Epub 2010 Sep 14, Fig 3. DOI 10.1242/?jcs.072900

    ab20346 used at a 1/1000 dilution in Western Blot.5-20µg of total protein from each sample was loaded.

  • Overlay histogram showing NIH3T3 cells stained with ab20346 (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 (ab20346, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in NIH3T3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

References for Anti-Vimentin antibody [VI-10] (ab20346)

This product has been referenced in:
  • van Marion MH  et al. Behavior of CMPCs in unidirectional constrained and stress-free 3D hydrogels. J Mol Cell Cardiol 87:79-91 (2015). IF ; Human . Read more (PubMed: 26278995) »
  • Zerr P  et al. Vitamin D receptor regulates TGF-ß signalling in systemic sclerosis. Ann Rheum Dis N/A:N/A (2014). Human . Read more (PubMed: 24448349) »

See all 25 Publications for this product

Product Wall

Application Immunocytochemistry
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
Sample Mouse Cultured Cells (thoracic aortic primary smooth muscle cells)
Specification thoracic aortic primary smooth muscle cells
Permeabilization Yes - 0.25% TritonX-100
Fixative Paraformaldehyde
Username

Mr. MC Shen

Verified customer

Submitted Dec 15 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (12,5)
Sample Human Tissue lysate - whole (human Urether)
Specification human Urether
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Fixation Frozen no other fixative
Permeabilization Yes - 0.1% Triton-X
Sample Human Cell (Brain)
Specification Brain
Gating Strategy forward scatter/side scatter (all cells)
Preparation Cell harvesting/tissue preparation method: Homogenized frozen tissue
Sample buffer: Tris/EDTA/sucrose/Triton
Username

Abcam user community

Verified customer

Submitted Sep 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample Human Cell (mammalian cancer cells)
Specification mammalian cancer cells
Permeabilization Yes - Tween20
Fixative Formaldehyde
Username

Mr. Gabriel Ortega

Verified customer

Submitted Jul 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 37°C
Sample Human Cell (Colon carcinoma)
Specification Colon carcinoma
Permeabilization Yes - 0.5% Triton in PBS
Fixative Paraformaldehyde
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Dec 04 2012

Thank you for contacting us. I am sorry to hear that the suggestions did not improve the performance of this product.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

Ho...

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Thank you for taking time to contact Abcam. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (mouse cardiac fibroblasts)
Loading amount 20 µg
Specification mouse cardiac fibroblasts
Gel Running Conditions Reduced Denaturing (10)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 02 2012

Thank you for contacting us.

The protocol we have for GMA staining is as follows;

Place biopsy immediately in ice cold acetone containing protease inhibitors
Fix overnight -20°C
Replace fixative with acetone (room tem...

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Thank you for your enquiry. I have contacted the originator of this antibody, and they have kindly provided the following information regarding the IP testing: 'The lysis was done by 1% lauryl maltoside + protease inhibitors. I am sorry we do not...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"