Anti-Vimentin antibody - Cytoskeleton Marker (ab45939)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab45939 in the following tested applications.

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

Application Notes
IHC-P 1/700. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).Can be blocked with Vimentin peptide (ab46156).
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr 1/50.

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.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • DomainThe central alpha-helical coiled-coil rod region mediates elementary homodimerization.
  • 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.
  • Cellular localizationCytoplasm.
  • Target information above from: UniProt accession P08670 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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 - Cytoskeleton Marker images

  • All lanes : Anti-Vimentin antibody - Cytoskeleton Marker (ab45939) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 4 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 54 kDa
    Observed band size : 54 kDa
  • All lanes : Anti-Vimentin antibody - Cytoskeleton Marker (ab45939) at 1 µg/ml

    Lane 1 : Human Vimentin full length protein (ab73843) at 0.1 µg
    Lane 2 : Human Vimentin full length protein (ab73843) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 54 kDa


    Exposure time : 10 seconds
  • ICC/IF image of ab45939 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45939, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Immunohistochemical detection (formaldehyde/paraffin-embedded sections) of vimentin protein using vimentin antibody - Neural Stem Cell Marker (ab45939) on human ovarian carcinoma sections. Antigen retrieval step:Heat mediated. Blocking step: 1% BSA for 10 mins at RT°C. Ab45939 was used at 1/700 for 1 hour. Secondary Antibody: Biotin-conjugated anti rabbit IG (1/300).
  • ab45939 staining Vimentin in human squamous cell carcinoma tissue sections by IHC-Fr (Frozen sections). Tissue samples were fixed with formaldehyde and blocked with 4% milk for 20 minutes at 22°C. The sample was incubated with primary antibody (1/50 in milk) at 4°C for 18 hours. A Biotin-conjugated Goat polyclonal to rabbit IgG (1/200) was used as secondary antibody.

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  • ab45939 staining Vimentin in murine kidney tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 10% serum for 30 minutes at room temperature followed by incubation with the primary antibody at a 1/2100 dilution for 18 hours at 4°C. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution.

    See Abreview

  • ICC/IF image of ab45939 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45939, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Vimentin antibody - Cytoskeleton Marker (ab45939)

This product has been referenced in:
  • Puschmann TB  et al. Bioactive 3D cell culture system minimizes cellular stress and maintains the in vivo-like morphological complexity of astroglial cells. Glia 61:432-40 (2013). WB ; Mouse . Read more (PubMed: 23292921) »
  • Wang L  et al. Impact of laminitis on the canonical Wnt signaling pathway in basal epithelial cells of the equine digital laminae. PLoS One 8:e56025 (2013). ICC/IF ; Horse . Read more (PubMed: 23405249) »

See all 14 Publications for this product

Product Wall

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (12% page)
Sample Human Cell lysate - whole cell (hep3b and L3.6PL)
Specification hep3b and L3.6PL
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Jan 16 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 4°C
Sample Human Cell (human ascites- pancreatic cancer cell line)
Specification human ascites- pancreatic cancer cell line
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 14 2014

Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guaran...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Mar 20 2012

Anbei wie versprochen die Publikationen aus der Gruppe von Gabbiani bzw. Bochaton-Piallat (PMIDs 21868702 bzw. 17347479). Ich hoffe, diese helfen Ihnen zumindest als Ausgangspunkt für Ihre weitere Suche nach geeigneten Markern. Vielleicht ist einer von...

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Application Western blot
Sample Human Recombinant protein (recombinant vimentin)
Loading amount 0.01 µg
Specification recombinant vimentin
Gel Running Conditions Reduced Denaturing
Blocking step Amersham ECL Blocking Agent as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted Aug 18 2011

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Kidney)
Specification Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Mike Forbes

Verified customer

Submitted Feb 09 2011

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (squamous cell carcinoma)
Specification squamous cell carcinoma
Fixative Formaldehyde
Permeabilization No
Blocking step Milk as blocking agent for 20 minute(s) · Concentration: 4% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Mar 01 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Monkey Tissue sections (Cerebellum)
Specification Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid ph6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Dec 21 2009

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Kidney and cerebellum)
Specification Kidney and cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid ph6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Mar 16 2009

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