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Anti-Vimentin antibody [V9] ab8069 is a mouse monoclonal antibody that is used in Vimentin western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and rat samples.

- Antibody clone V9 is the most widely used clone for Vimentin on the market
- Specificity confirmed with VIM knockout cell line validation
- One antibody for all your Vimentin staining


Images

Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB8069), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB8069), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB8069), expandable thumbnail
  • Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB8069), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB8069), expandable thumbnail

Publications

Key facts

Isotype
IgG1
Host species
Mouse
Storage buffer

pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PFlow Cyt (Intra)WBICC/IF
Human
Tested
Tested
Tested
Tested
Rat
Expected
Tested
Expected
Tested
Cat
Predicted
Predicted
Predicted
Predicted
Chicken
Predicted
Predicted
Predicted
Predicted
Cow
Predicted
Predicted
Predicted
Predicted
Dog
Predicted
Predicted
Predicted
Predicted
Horse
Predicted
Predicted
Predicted
Predicted
Pig
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species
Human
Dilution info
0.50000-5.00000 µg/mL
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Horse, Chicken, Cow, Cat, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Rat
Dilution info
0.10000-1.00000 µg for 106 Cells
Notes

Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Species
Human
Dilution info
0.10000-1.00000 µg for 106 Cells
Notes

Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Predicted
Predicted

Species
Horse, Chicken, Cow, Cat, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1 µg/mL
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Horse, Chicken, Cow, Cat, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Rat
Dilution info
0.50000-5.00000 µg/mL
Notes

-

Species
Human
Dilution info
0.50000-5.00000 µg/mL
Notes

-

Predicted
Predicted

Species
Horse, Chicken, Cow, Cat, Dog, Pig
Dilution info
-
Notes

-

Associated Products

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Target data

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. Plays a role in cell directional movement, orientation, cell sheet organization and Golgi complex polarization at the cell migration front (By similarity). Protects SCRIB from proteasomal degradation and facilitates its localization to intermediate filaments in a cell contact-mediated manner (By similarity). Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.

Alternative names

Recommended products

Anti-Vimentin antibody [V9] ab8069 is a mouse monoclonal antibody that is used in Vimentin western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and rat samples.

- Antibody clone V9 is the most widely used clone for Vimentin on the market
- Specificity confirmed with VIM knockout cell line validation
- One antibody for all your Vimentin staining

Key facts

Isotype
IgG1
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
V9
Purification technique
Affinity purification Protein G
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

This monoclonal antibody to vimentin has been knockout validated in ICC/IF. The expected staining was observed in wild type cells and no staining was seen in vimentin knockout cells.

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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14 product images

  • Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail
    Image from Tange S et al., PLoS One. 2014;9(12):e115684. Fig 9(H).; doi: 10.1371/journal.pone.0115684. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    All lanes: Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Lane 1: Whole cell lysate of A549 cells

    Lane 2: Whole cell lysate of A549 cells treated with TGF-beta

    Lane 3: Whole cell lysate of A549 cells overexpressing JARID2

    Lane 4: Whole cell lysate of A549 cells overexpressing JARID2 treated with TGF-beta

    Predicted band size: 53 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail
    Image from Loessner D et al, Biomaterials. 2010 Nov;31(32):8494-506. Epub 2010 Aug 14. doi:10.1016/j.biomaterials.2010.07.064

    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    ab8069 staining Vimentin in human epithelial ovarian serous adenocarcinoma cell line SKOV-3 by Immunocytochemistry/ Immunofluorescence.

    Samples were fixed with 4% PFA in PBS pH 7.4 and then permeabilised using 0.2% saponin for 30 minutes. A blocking step was performed using 1% BSA/PBS for 1 hour. Samples were then incubated with ab8069 at a 1/200 dilution in 1% BSA/PBS for 1 hour. The secondary antibody was a goat anti-mouse Alexa 488 (green) diluted 1/1000, 1% BSA/PBS for 1 hour. Samples were then incubated with phalloidin (red for actin staining) in 1% BSA/PBS for 45 minutes and counterstained with DAPI (blue for nuclei staining) in PBS for 45 minutes.

  • Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Flow cytometry overlay histogram showing wild-type HAP1 (green line) and VIM knockout HAP1 cells stained with ab8069 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab8069) (1x106 in 100 µl at 0.04 µg/ml) for 30 min at 22°C.

    The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 22°C.

    Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line VIM knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a .

    This antibody gave a positive signal in VIM HAP1 knockout cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions./

  • Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Lanes 1 - 4: Merged signal (red and green). Green - ab8069 observed at 53 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 observed at 37 kDa.

    ab8069 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human VIM (Vimentin) knockout HeLa cell line ab255446 (knockout cell lysate Human VIM (Vimentin) knockout HeLa cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab8069 and Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) were incubated overnight at 4°C at 1 μg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069) at 1 µg/mL

    Lane 1: U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate at 20 µg

    Lane 2: Human tonsil cell lysate at 20 µg

    Lane 2: Western blot - Human VIM (Vimentin) knockout HeLa cell line (Human VIM (Vimentin) knockout HeLa cell line ab255446)

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: VIM knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 53 kDa

    Observed band size: 50 kDa, 53 kDa

  • Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Overlay histogram showing SV40LT-SMC cells stained with ab8069 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab8069, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in SV40LT-SMC cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    ab8069 staining Vimentin (colored green) in wild-type HAP1 cells (top panel) and VIM 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 ab8069 at 0.5μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272 (Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 594)) at 1/250 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 (Goat secondary antibody to Mouse IgG (Alexa Fluor® 488)) at 2 μg/ml (colored green). Nuclear DNA was labelled in blue with DAPI.

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

  • Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Overlay histogram showing MDA-MB-231 cells stained with ab8069 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab8069, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in MDA-MB-231 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    Lanes 1 - 4: Merged signal (red and green). Green - ab8069 observed at 57 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.

    ab8069 was shown to specifically react with Vimentin in wild-type HAP1 cells as signal was lost in VIM (Vimentin) knockout cells. Wild-type and VIM (Vimentin) knockout samples were subjected to SDS-PAGE. ab8069 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069) at 1 µg/mL

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: VIM (Vimentin) knockout HAP1 whole cell lysate at 20 µg

    Lane 3: HeLa whole cell lysate at 20 µg

    Lane 4: MOLT4 whole cell lysate at 20 µg

    Predicted band size: 53 kDa

  • Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    All lanes: Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069) at 1 µg/mL

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate at 10 µg

    Lane 2: MOLT4 (Human lymphoblastic leukemia cell line) Whole Cell Lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 53 kDa

    Observed band size: 57 kDa

    Exposure time: 20min

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    ab8069 staining Vimentin in HeLa cells. The cells were fixed with 4% formaldehyde (10min), 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 overnight at +4°C with ab8069 at 1/100 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2μg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    ab8069 staining Vimentin in rat glial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 10 hours at 4°C. An Alexa Fluor® 555-conjugated anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    ab8069 staining Vimentin in Human oral cavity tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde and blocked with Protein Block ab64226 Protein Block for 5 minutes at 25°C; antigen retrieval was by heat mediation in pH 6 buffer . Samples were incubated with primary antibody (1/1000 in 10% NGS) for 16 hours at 4°C. An undiluted biotinylated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    IHC image showing no staining when ab8069 was used on mousekidney formalin fixed paraffin embedded tissue, using MOM detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab8069 at 5 μg/ml for 15 mins at room temperature. DAB was used as the chromogen. The section was 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069), expandable thumbnail
    Lab

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)

    IHC image of Vimentin staining in a section of formalin-fixed paraffin-embedded human normal kidney* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab8069, 0.5ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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