Anti-Vimentin antibody [SP20] (ab16700)

Knockout Tested Rabbit monoclonal Vimentin antibody [SP20]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 33 publication(s). Independently reviewed in 6 review(s).


  • Product name
    Anti-Vimentin antibody [SP20]
    See all Vimentin primary antibodies
  • Description
    Rabbit monoclonal [SP20] to Vimentin
  • Host species
  • Specificity
    We have data to show that ab16700 is not suitable for work on mouse tissue. For researchers working on mouse we recommend using ab92547. If you would like further information on this, please do not hesitate to contact our technical support team.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Fr, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat, Hamster, Cow, Xenopus laevis
  • Immunogen

    Recombinant full length protein within Human Vimentin. The exact sequence is proprietary.

  • Positive control
    • Sarcomas, HeLa whole cell lysate (ab150035) ICC/IF: HAP1-VIM cells



Our Abpromise guarantee covers the use of ab16700 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 1/1000.
WB Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-Fr Use at an assay dependent concentration. PubMed: 19554024
IHC-P 1/200.

Antigen Retrieval: None

Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.


  • Function
    Vimentins 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 specificity
    Highly 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 disease
    Cataract 30
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Domain
    The 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
    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 localization
  • Information by UniProt
  • Database links
  • Form
    Vimentin 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 [SP20] (ab16700) at 1/100 dilution + HeLa cell lysate

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

  • ab16700 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab16700 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemical analysis of Human melanona tissue labelling Vimentin with ab16700.

  • Immunohistochemical analysis of Human colorectal cancer tissue, staining Vimentin (red) with ab16700.

    Tissue was fixed with ice cold acetone for 5 minutes and blocked with 10% goat serum for 30 minutes. Samples were incubated with primary antibody (1/20) and a Cy5&reg-conjugated anti-rabbit IgG (1/200) was used as the secondary antibody.

  • IHC image of ab16700 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16700, (Neat supernatant) for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab16700 staining Vimentin in human corneal limbal epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.3% Triton X-100 for 5 minutes. Samples were incubated with primary antibody (1/200 in PBS + 10% normal goat serum) for 18 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • ab16700 at 1/200 staining Human Limbal Epithelial Cells by ICC/IF. The cells were incubated with the antibody for 1 hour and then a FITC conjugated goat antibody was used as the secondary. The image shows vimentin staining in green and hoechst staining in blue. The upper cells in the image (vimentin negative) are epithelium cells. the vimentin positive cells are stroma cells.

    See Abreview

  • Flow cytometric analysis of rabbit anti-Vimentin (SP20) antibody ab16700 (1/100) in HeLa cells (green) compared to negative control of rabbit IgG (blue).

  • Overlay histogram showing HeLa cells stained with ab16700 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab16700, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.


This product has been referenced in:
  • Pakula M  et al. The Epithelial-Mesenchymal Transition Initiated by Malignant Ascites Underlies the Transmesothelial Invasion of Ovarian Cancer Cells. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30609691) »
  • Biskou O  et al. The type III intermediate filament vimentin regulates organelle distribution and modulates autophagy. PLoS One 14:e0209665 (2019). Read more (PubMed: 30699149) »
See all 40 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence
Human Cell (MDA-MB-231)
Yes - 0.5% Triton X for 15 min
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Jorge Guerrero

Verified customer

Submitted Jun 19 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Rabbit Tissue sections (Corneal limbus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA (ph 9.0)
Yes - PBT: 0.1% Triton X-100 in PBS
Corneal limbus
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Noelia Andollo

Verified customer

Submitted Oct 08 2018

Immunohistochemistry (Frozen sections)
Mouse Tissue sections (Testis, adult)
Yes - 0.1% Triton X-100 in PBS
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Mr. Bryan Niedenberger

Verified customer

Submitted Jul 12 2016

Western blot
Dog Cell lysate - whole cell (Osteosarcoma cell lines)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Osteosarcoma cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Holly Pondenis

Verified customer

Submitted Nov 21 2015

Immunohistochemistry (Frozen sections)
Mouse Tissue sections (Pancreas)
Blocking step
CAS block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 09 2014

Immunocytochemistry/ Immunofluorescence
Human Cell (Corneal limbal epithelial cells)
Corneal limbal epithelial cells
Yes - Triton 0,3% - 5 minutes

Abcam user community

Verified customer

Submitted Jan 13 2014

Immunocytochemistry/ Immunofluorescence
Human Cell (Human Limbal Stromal cells)
Human Limbal Stromal cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2%

Miss. Ashley Quintana

Verified customer

Submitted Nov 20 2007

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