Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (AB110252)

ab110252 stained HCT116 cells. The cells were 100% methanol fixed for 5 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110252 at 1/50 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• Flow Cyt

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Flow Cytometry - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (AB110252)

Overlay histogram showing HepG2 cells stained with ab110252 (red line). The cells were fixed with 80% methanol (5 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. The cells were then incubated with the antibody (ab110252, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• WB

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Western blot - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (AB110252)

Extra bands in the Mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.

All lanes:

Western blot - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (ab110252) at 0.5 µg/mL

Lane 1:

Human heart mitochondrial lysate

Lane 2:

Cow heart mitochondrial lysate

Lane 3:

Rat heart mitochondrial lysate

Lane 4:

Mouse heart mitochondrial lysate

Lane 5:

HepG2 mitochondrial lysate

Predicted band size: 53 kDa

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