Anti-Vimentin antibody [VI-RE/1]

6 product images

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  Flow Cytometry (Intracellular) - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  Flow cytometry (Intracellular) analysis ESS-1 (human endometrial stromal sarcoma cell line) cells labeling Vimentin with ab3974 (GAM-FITC).
  

  Negative control: human lymphocytes.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  Immunohistochemistry staining (paraffin sections) of Vimentin in human liver using mouse monoclonal antibody [VI-RE/1] (ab3974, diluted 1/400), detected with GAM IgG-Alexa Fluor®488 (diluted 1/200; green), cell nuclei stained with PI (1 μg/ml; orange).

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  Immunocytochemistry - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  ICC image of ab3974 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 (ab3974, 5μg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879, DyLight® 488 goat anti-mouse IgG (H+L) 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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  IHC image of Vimentin staining in human bladder formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab3974, 5μg/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.

  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.

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  Flow Cytometry (Intracellular) - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  Flow Cytometry (Intracellular) analysis of HeLa cells labeling Vimentin with ab3974 (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 (ab3974, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 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, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  

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  Western blot - Anti-Vimentin antibody [VI-RE/1] (ab3974)

  All lanes: Western blot - Anti-Vimentin antibody [VI-RE/1] (ab3974) at 5 µg/mL

  All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Predicted band size: 53 kDa

  Observed band size: 57 kDa