All lanes : Anti-ATP5C1 antibody [2A1AA11] (ab119686) at 1 µg/ml

Lane 1 : human heart homogenate lysate at 15 µg
Lane 2 : human HepG2 cell lysate at 15 µg
Lane 3 : human liver mitochondria lysate at 7.5 µg
Lane 4 : rat liver mitochondria lysate at 7.5 µg
Lane 5 : mouse liver mitochondria lysate at 7.5 µg
Lane 6 : bovine heart mitochondria lysate at 7.5 µg

Secondary
All lanes : Goat anti-mouse HRP at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 33 kDa

Immunocytochemistry using ab119686 stained HDFn cells (human). The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min) with antigen retrieval. The cells were then incubated with the antibody (ab119686, 1µg/ml) for 2h at room temperature or over night at 4°C. The secondary antibody was (red) 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondria.

Flow cytometry. Hela cells were stained with 1µg/mL anti-ATP5C1 antibody (ab119686) (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

IHC image of ATP5C1 staining in Human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab119686, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.