Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker (AB196847)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 μg/ml PHA for 18h (bottom), either stained with ab196847 (right) or isotype control (left). The cells were fixed using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab196847) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker (AB196847)

ab196847 staining AIF in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 10% normal goat serum in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196847 at 1/2500 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/200 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker (AB196847)

ab196847 staining AIF in HACAT cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 10% normal goat serum in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196847 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/200 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).