FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (AB119688)

Flow cytometry overlay histogram showing HeLa cells stained with ab119688 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 90% methanol for 30 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab119688) (1x 10⁶ in 100μl at 1.0μg/ml) for 30min at 22°C.

Isotype control antibody (black line) was ab314952 (1.0μg/ml (1/500)) 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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (AB119688)

Overlay histogram showing HeLa cells stained with ab119688 (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 / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab119688, 1/100 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was mouse IgG2b FITC (ab18419) 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 50 mW Blue laser (488nm) and 530/30 bandpass filter.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (AB119688)

Mitochondrial localization of ATP5A using antibody ab119688. Cultured HeLa cells were fixed, permeabilized and then labeled with 15H4C4-FITC (1 µg/ml). Since the antibody is labeled with FITC no secondary antibody is necessary.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (AB119688)

ab119688 staining ATP5A in HeLa 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 ab119688 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (pseudocolored 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 100% methanol (5min).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - FITC Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (AB119688)

Overlay histogram showing HeLa cells stained with ab119688 (blue line). The cells were fixed with 4% paraformaldehyde (10 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 ab119688 (15H4C4 FITC) at 1μg/1xE6/mL cells for 30 min at 22°C. Negative control (red line) is unstained d cells. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.