Anti-MTCO2 antibody [EPR3314] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MTCO2 antibody [EPR3314] - BSA and Azide free (AB239889)

Overlay histogram showing HepG2 cells stained with ab79393 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab79393, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79393).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MTCO2 antibody [EPR3314] - BSA and Azide free (AB239889)

ICC/IF image of ab79393 stained HepG2 cells. The cells were 100% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79393, 5μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79393).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO2 antibody [EPR3314] - BSA and Azide free (AB239889)

ab79393 at 1/100 dilution staining Cytochrome C oxidase subunit II in human liver (A), human heart (B) and HeLa cell line (C) by Immunohistochemistry, Paraffin-embedded tissues. Detection method of A and B was HRP-conjugated anti-rabbit with DAB substrate used for staining.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79393).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-MTCO2 antibody [EPR3314] - BSA and Azide free (AB239889)

This data was developed using ab79393, the same antibody clone in a different buffer formulation.
Purified ab79393 at 1/50 dilution (2μg) immunoprecipitating MTCO2 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab79393 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab79393 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 21 kDa

All lanes:

Immunoprecipitation - Anti-MTCO2 antibody [EPR3314] (<a href='/en-us/products/primary-antibodies/mtco2-antibody-epr3314-ab79393'>ab79393</a>)

Predicted band size: 25 kDa

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