Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker

3 product images

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  Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab195877)

  Overlay histogram showing HeLa cells stained with ab195877 (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 (ab195877, 1/500 dilution) for 30 min at 22°C.
  

  Isotype control antibody (black line) was mouse IgG1 (monoclonal) Alexa Fluor® 488 (ab171463) used at the same concentration and conditions as the primary antibody. 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 HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

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  Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab195877)

  ab195877 staining Vimentin (shown in green) 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 ab195877 at 1/1000 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
  

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

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  Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab195877)

  ab195877 staining Vimentin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab195877 at 1/1000 dilution (shown in green) and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin at 1μg/ml overnight at +4°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed, was then incubated at 2μg/ml for 1h at room temperature (shown in red) Nuclear DNA was labelled in blue with DAPI.

  This product gave a positive signal in 100% methanol (5 min) fixed HeLa cells under the same testing conditions.

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