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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB195877)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Vimentin with ab195877 at 1/100 (5 μg/ml).

Positive staining on human breast cancer.

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Vimentin antibody [V9] - Cytoskeleton Marker (AB195877)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Vimentin with ab195877 at 1/100 (5 μg/ml).

Positive staining on human kidney.

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• Flow Cyt (Intra)

Unknown

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.

• ICC/IF

Lab

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 ab6046, Rabbit polyclonal to beta Tubulin at 1μg/ml overnight at +4°C. 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).

• ICC/IF

Lab

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 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).