Alexa Fluor® 405 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 405 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (AB210152)

Overlay histogram showing HeLa cells stained with ab210152 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min, blocked with Image-iT® FX Signal Enhancer for 30 min at 22°C and then blocked with 1x PBS / 10% normal goat serum for 1 hr at 22°C. Cells were then incubated with the antibody (ab210152, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 405 (ab208150) 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 50mW violet laser (405nm) and 450/50 bandpass filter.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 405 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (AB210152)

Overlay histogram showing HAP1 wildtype (green line) and HAP1-VIM knockout cells (red line) stained with ab210152. The cells were fixed with 4% formaldehyde (10 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 (ab210152, 0.1μg/1x106 cells OR 1/500 dilution) for 30 min at 22°C.

A rabbit IgG isotype control antibody (ab208150) 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 Violet laser (405nm) and 450/50 bandpass filter.

This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.