Anti-SDHB 抗体 [21A11AE7]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SDHB antibody [21A11AE7] (AB14714)

ab14714 staining SDHB in wild-type HEK293 cells (top panel) and SDHB knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab14714 at 5μg concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt

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Flow Cytometry - Anti-SDHB antibody [21A11AE7] (AB14714)

Overlay histogram showing HEK-293 (human epithelial cell line from embryonic kidney) 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SDHB antibody [21A11AE7] (AB14714)

ICC/IF image of ab14714 stained HeLa (human epithelial cell line from cervix adenocarcinoma) 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.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-SDHB antibody [21A11AE7] (AB14714)

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SDHB antibody [21A11AE7] (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.

• WB

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Western blot - Anti-SDHB antibody [21A11AE7] (AB14714)

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.

All lanes:

Western blot - Anti-SDHB antibody [21A11AE7] (ab14714) at 5 µg/mL

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 (human liver hepatocellular carcinoma cell line) cells at 20 µg

Secondary

All lanes:

Goat anti-Mouse secondary

Predicted band size: 32 kDa

Observed band size: 28 kDa,55 kDa

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