Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Flow cytometry overlay histogram of 4% paraformaldehyde HEK-293 (human embryonic kidney epithelial cell) cells permeabilised with 90% methanol labelling Vinculin with ab196454 at 1/500 dilution (Red line) compared with a Rabbit monoclonal IgG (ab172730) as Isotype control (Black line) and an unlabelled control (cells without incubation with primary antibody) (Blue line).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Flow cytometry overlay histogram showing left HeLa positive cells and right negative HeLa VCL KO stained with ab196454 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab196454) (1x 10⁶ in 100μl at 0.008μg/ml (1/62500)) for 30min at 22°C.

Isotype control antibody (black line) was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

ab196454 staining Vinculin in HUVEC cells. The cells were fixed with 4% formaldehyde (10 min), 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 ab196454 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 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 HUVEC cells fixed with 100% methanol (5min).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Flow cytometry overlay histogram of 4% paraformaldehyde C6 (rat glial tumor glial cell) cells permeabilised with 90% methanol labelling Vinculin with ab196454 at 1/500 dilution (Red line) compared with a Rabbit monoclonal IgG (ab172730) as Isotype control (Black line) and an unlabelled control (cells without incubation with primary antibody) (Blue line).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Immunofluorescent analysis of 100% methanol fixed NIH/3T3 (mouse embryonic fibroblasts) permeabilized using 0.1% Triton X-100 labelling Vinculin with ab196454 at 1/50 dilution (green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (magenta) was used as a counterstain. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in NIH/3T3 cell line was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Immunofluorescent analysis of 100% methanol fixed C6 (rat glial tumor glial cell) permeabilized using 0.1% Triton X-100 labelling Vinculin with ab196454 used at 1/50 dilution (green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (magenta) was used as a counterstain. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in C6 cell line was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Flow cytometry overlay histogram of 4% paraformaldehyde NIH/3T3 (mouse embryonic fibroblast) cells permeabilised with 90% methanol labelling Vinculin with ab196454 at 1/500 dilution (Red line) compared with a Rabbit monoclonal IgG (ab172730) as Isotype control (Black line) and an unlabelled control (cells without incubation with primary antibody) (Blue line).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (AB196454)

Flow cytometry overlay histogram of 4% paraformaldehyde HUVEC (human umbilical vein endothelial cell) cells permeabilised with 90% methanol labelling Vinculin with ab196454 at 1/500 dilution (Red line) compared with a Rabbit monoclonal IgG (ab172730) as Isotype control (Black line) and an unlabelled control (cells without incubation with primary antibody) (Blue line).