All lanes : Anti-ATP5F1 antibody [9D1BC4] (ab117991) at 1 µg/ml

Lane 1 : Human heart homogenate at 15 µg
Lane 2 : HepG2 lysate at 15 µg
Lane 3 : Human liver mitochondria at 7.5 µg
Lane 4 : Bovine heart mitochondria at 7.5 µg
Lane 5 : Rat liver mitochondria at 7.5 µg
Lane 6 : Mouse liver mitochondria at 7.5 µg

Predicted band size: 24.6 kDa



Note - Extra bands seen in lane 6 are secondary antibody mouse-on-mouse effects and not related to the primary antibody.

ab117991 at 1 ug/ml with Hela cells in flow cytometry.

Immunocytochemistry image of ab117991 (MS972) stained NIH3T3 cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min) with urea/heat antigen retrieval method. The cells were incubated with ab117991 at 1 µg/ml for 2h at room temperature or over night at 4°C. The secondary antibody was (red) AlexaFluor® 594goat 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. DAPI was used to stain the cell nuclei (blue). The target protein locates to the mitochondria. The four cells in the upper portion of the image show mitochondria in the elongated, reticular, arrangement, the three cells in the lower portion of the image show a punctuate mitochondrial organization and may be dividing/have recently divided.

IHC image of ATP5F1 staining in Human normal colon 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 ab117991, 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.