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
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
IHC-P | Flow Cyt (Intra) | WB | ICC/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 |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.50000-5.00000 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Horse, Chicken, Cow, Cat, Dog, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Horse, Chicken, Cow, Cat, Dog, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Horse, Chicken, Cow, Cat, Dog, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 0.50000-5.00000 µg/mL | Notes - |
Species Human | Dilution info 0.50000-5.00000 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Horse, Chicken, Cow, Cat, Dog, Pig | Dilution info - | Notes - |
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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.
Vimentin, VIM
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
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
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.
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This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
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
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 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./
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
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
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.
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).
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.
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
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
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
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).
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.
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.
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.
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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