All lanes : Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684) at 1 µg/ml

Lane 1 : recombinant mtTFA (ab103784) at 0.025 µg
Lane 2 : HeLa whole cell lysate at 30 µg

Secondary
All lanes : HRP-conjugated Goat polyclonal to mouse IgG at 1/5000 dilution

Predicted band size: 29 kDa

Immunocytochemistry image of ab119684 stained human HDFn cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min). The cells were incubated with the antibody (18G102B2E11, 1µg/ml) for 2h at room temperature. The secondary antibody was (green) 488 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. DAPI was used to stain the cell nuclei (blue). mtTFA localizes to the mitochondria.

HeLa cells were stained with 2 µg/mL mtTFA antibody (ab119684) (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

IHC image of mtTFA staining in human liver HCC formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab119684, 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.