Alexa Fluor® 647 Anti-Fibronectin antibody [F14]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Fibronectin antibody [F14] (AB246736)

ab246736 staining Fibronectin in HepG2 cells. The cells were fixed with 100% methanol (5 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 ab246736 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). 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 HepG2 cells fixed with 4% formaldehyde (10 min).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Fibronectin antibody [F14] (AB246736)

Overlay histogram showing HepG2 cells stained with ab246736 (red line). 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 containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab246736) (1x 10⁶ in 100μl at 0.2μg/ml (1/2500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor^® 647 (ab199093) 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 40 mW Red laser (640nm) and 670/14 bandpass filter.

This antibody gave a positive signal inHepG2 cells fixed with 4 % formaldehyde (10 min) / permeabilized with 0.1 % PBS-Triton X-100 for 15 min used under the same conditions.