Immunocytochemistry image of ab110293 stained Human HepG2 cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min). The cells were incubated with ab110293 at 4 µg/ml for 2h at room temperature, or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 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. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.

HL-60 cells were stained with 1 µg/mL of ab110293 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

SDS-PAGE analysis of IP samples. As an IP antibody, ab110293 works with human, bovine, rat, and mouse samples.

Lane STD: Molecular weight ladder

Lane HLM: Human liver mitochondria

Lane BHM: Bovine heart mitochondria

Lane RLM: Rat liver mitochondria

Lane MLM: Mouse liver mitochondria

IHC image of CPT2 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 ab110293, 10µg/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.