Anti-SDHB antibody [21A11AE7] (ab14714)

  Western blot - SDHB antibody [21A11] (ab14714)

All lanes : Anti-SDHB antibody [21A11AE7] (ab14714)

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 10 µg
Lane 5 : Isolated mitochondria from HepG2 cells at 20 µg

Secondary
Goat anti-Mouse secondary

Observed band size : 28 kDa (why is the actual band size different from the predicted?)
Additional bands at : 55 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 (Formalin/PFA-fixed paraffin-embedded sections) - SDHB antibody [21A11] (ab14714)

ab14714 at 5ug/ml staining human adrenal tumor tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. A biotinylated horse anti-mouse IgG antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

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  Immunocytochemistry/ Immunofluorescence - SDHB antibody [21A11] (ab14714)

ICC/IF image of ab14714 stained HeLa cells. The cells were 4% formaldehyde 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 (ab14714, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  Immunocytochemistry/ Immunofluorescence - SDHB antibody [21A11] (ab14714)

Mitochondrial localization of complex II visualized by immunocytochemistry using anti-complex II subunit 30 kDa Ip mAb 21A11 (ab14714). Cells were fixed, permeabilized and then labeled with ab14714 followed by an Alexa Fluor® 488-conjugated-goat-anti-mouse IgG2a isotype specific secondary antibody.

  Immunohistochemistry (Frozen sections) - SDHB antibody [21A11] (ab14714)

ab14714 staining SDHB in normal ageing human colon tissue by Immunohistochemistry (Frozen sections).

  Flow Cytometry-SDHB antibody [21A11](ab14714)

Overlay histogram showing HEK293 cells stained with ab14714 (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 (ab14714, 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 IgG2a [ICIGG2A] (ab91361, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.