Anti-MTCO1 antibody [1D6E1A8] - Mitochondrial Marker (ab14705)

Immunocytochemistry/ Immunofluorescence - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

ab14705 staining MTCO1 - Mitochondrial Marker in pig retinal pigment epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/2000 in 1% goat serum, 0.1% TX100; PBS) for 16 hours at 4°C. An Alexa Fluor^®546-conjugated Goat polyclonal to mouse IgG, dilution 1/500, was used as secondary antibody.

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

Immunocytochemistry/ Immunofluorescence - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

ab14705 staining MTCO1 - Mitochondrial Marker in rat cerebellum primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/2000 in 1% goat serum, 0.1% TX100; PBS) for 16 hours at 4°C. An Alexa Fluor^®546-conjugated Goat polyclonal to mouse IgG, dilution 1/5000, was used as secondary antibody.

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

Flow Cytometry - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

Overlay histogram showing HEK293 cells stained with ab14705 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab14705, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

Western blot - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

All lanes : Anti-MTCO1 antibody [1D6E1A8] - Mitochondrial Marker (ab14705)

Lane 1 : Isolated mitochondria from Human heart at 5 µg
Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
Lane 3 : Isolated mitochondria from Rat heart at 10 µg
Lane 4 : Isolated mitochondria from Mouse heart at 5 µg

Secondary
Goat Anti-Mouse IgG

Predicted band size : 57 kDa
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.

Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.

Immunohistochemistry (Frozen sections) - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

ab14705 staining MTCO1 in skeletal muscle tissue by Immunohistochemistry (Frozen sections). Tissue sections were from from a patient with a single large deletion of the mtDNA and show a mosaic of complex IV positive and complex IV negative fibers.

Immunohistochemistry (Frozen sections) - MTCO1 antibody [1D6] - Mitochondrial Marker (ab14705)

ab14705 staining MTCO1 in Human colon tissue by Immunohistochemistry (Frozen sections). Tissue sections from a normal ageing patient show complex IV negative crypts due to clonal expansion of colonic stem cells bearing mutations in the mtDNA-encoded gene for complex IV.