All lanes : Anti-ACADM/MCAD antibody [3B7BH7] (ab110296) at 0.125 µg/ml

Lane 1 : HepG2 cell lysates
Lane 2 : HeLa cell lysates
Lane 3 : H9C2 (rat cells) lysates
Lane 4 : MEF (mouse cells) lysates

Lysates/proteins at 15 µg per lane.

Predicted band size: 47 kDa

Immunocytochemistry image of ab110296 stained Human HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with ab110296 (1 µg/ml) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (green) Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. 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.

ACADM/MCAD immunohistochemistry in Human cerebellum visualized with ab110296 at 1/1000. MCAD immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells. Note the distinctive subcellular localization of MCAD immunoreactivity in the Purkinje cell bodies.

HL-60 cells were stained with 1 µg/mL ab110296 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

HL-60 cells were incubated at 37&degC for 24h with vehicle control (0 &microM) and different concentrations of fenofibrate (ab120832). Increased expression of ACADM/MCAD in HL-60 cells correlates with an increase in fenofibrate concentration, as described in literature.

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab110296 at 1 &microg/ml and ab9484 at 1 &microg/ml overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (ab97040) at 1/10000 dilution and visualised using ECL development solution.