All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml

Lane 1 : Human skeletal muscle tissue lysate
Lane 2 : Rat skeletal muscle tissue

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Blocking buffer: 2% BSA

Gel type: MES

Exposure Time: 1 minute

Lane 1 : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml (Human skeletal muscle tissue lysate)
Lanes 2-3 : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml

Lane 2 : Human heart tissue lysate
Lane 3 : Mouse skeletal muscle tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Blocking buffer: 2% BSA

Gel type: MES

Exposure Time: 1 minute

All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml

Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Human liver tissue lysate - total protein (ab29889)
Lane 3 : Human heart tissue lysate - total protein (ab29431)
Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 17 kDa

ab16056 staining COX IV in Mouse heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/600 ) for 12 hours at 4°C. A Cy5® donkey anti-rabbit secondary antibody was used as the secondary antibody.

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ab16056 stained in Hela cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab16056 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 0.38 µg/ml

Lane 1 : HeLa whole cell lysate
Lane 2 : Human skeletal muscle cell lysate
Lane 3 : HeLa whole cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml
Lane 4 : Human skeletal muscle cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

IHC image of COXIV staining in human liver FFPE section, performed on a 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 ab16056, 1µg/ml, for 8 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.

ab16056 staining COX IV in breast tumour tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% buffered formalin and blocked with 5% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a 10mM sodium citrate buffer pH6. Samples were incubated with primary antibody (1/300 in blocking buffer) for 12 hours at 4°C. An Alexa Fluor^® 647-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

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