PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker

3 product images

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  Flow Cytometry (Intracellular) - PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab209446)

  Overlay histogram showing HAP1 wildtype (green line) and HAP1-VIM knockout cells (red line) stained with ab209446. 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 / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209446, 0.01μg/1x106 cells OR 1/5000 dilution) for 30 min at 22°C.A rabbit IgG isotype control antibody (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-VIM knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab209446)

  ab209446 staining Vimentin in HeLa cells. 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 overnight at +4°C with ab209446 at 1/500 dilution (Pseudocolored in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
  

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

  This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

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  Flow Cytometry (Intracellular) - PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab209446)

  Overlay histogram showing HeLa cells stained with ab209446 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol (-20°C) for 30 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209446, 1/5000 dilution) for 30 min at 22°C.
  

  Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycorythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) 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 20 mW Solid State Blue Laser (488nm) and 585/40 bandpass filter.