Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745)

  Western blot - UQCRC2 antibody [13G12] (ab14745)

All lanes : Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745) at 0.5 µg/ml

Lane 1 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)
Lane 2 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Observed band size : 48 kDa (why is the actual band size different from the predicted?)
Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - UQCRC2 antibody [13G12] (ab14745)

IHC image of ab14745 staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab14745, 1µg/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.

  Flow Cytometry-UQCRC2 antibody [13G12](ab14745)

Overlay histogram showing HepG2 cells stained with ab14745 (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 (ab14745, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

  - Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745)

Mitochondrial localization of complex III visualized by immunocytochemistry using anti-complex III subunit Core 2 mAb 2E3GC12FB2AE2 (MS304). Cultured human embryonic lung-derived fibroblasts (strain MRC5) were fixed, permeabilized and then labeled with MS304 (0.5 µg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 µg/ml).