Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432) at 1 µg/ml + Human testis tissue lysate - total protein (ab30257) at 20 µg

Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 57 kDa
Observed band size: 57 kDa


Exposure time: 16 minutes

Immunocytochemistry/Immunofluorescent analysis of ATPB (red) in HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with ab5432 at 1:100 overnight at 4ºC in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification.

Immunofluorescent analysis of ATPB in A431 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a ATPB monoclonal antibody (ab5432) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. ATPB staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunofluorescent analysis of ATPB in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a ATPB monoclonal antibody (ab5432) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. ATPB staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunoprecipatation of rat neuronal/glial cell extract using ab5432.

ICC/IF image of ab5432 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5432, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunocytochemistry/Immunofluorescence analysis of rat neuronal/glial cell culture using ab5432.