Anti-BCKDHA antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BCKDHA antibody (AB126173)

Immunofluorescence analysis of HeLa cells fixed in ice-cold MeOH for 5 min labelling BCKDHA protein at mitochondria using ab126173 at a 1/400 dilution (Green). Hoechst 33342 was used for nuclear staining (Blue).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA antibody (AB126173)

ab126173, at 1/250 dilution, staining BCKDHA in paraffin-embedded Human Colon cancer tissue by Immunohistochemistry.

• WB

Lab

Western blot - Anti-BCKDHA antibody (AB126173)

False colour image of Western blot : Anti-BCKDHA antibody staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab126173 was shown to bind specifically to BCKDHA. A band was observed at 48 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BCKDHA knockout cell line ab266439 (knockout cell lysate ab258324). To generate this image, wild-type and BCKDHA knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-BCKDHA antibody (ab126173) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

BCKDHA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human BCKDHA knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-bckdha-knockout-hek-293t-cell-line-ab266439'>ab266439</a>)

Lane 3:

Human Heart cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 48 kDa

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• WB

Supplier Data

Western blot - Anti-BCKDHA antibody (AB126173)

Samples were separated by 10% SDS PAGE.

All lanes:

Western blot - Anti-BCKDHA antibody (ab126173) at 1/1000 dilution

All lanes:

HeLa Whole cell extract at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-BCKDHA antibody (AB126173)

10% SDS PAGE

All lanes:

Western blot - Anti-BCKDHA antibody (ab126173) at 1/1000 dilution

All lanes:

HepG2 whole cell lysate at 30 µg

Predicted band size: 50 kDa

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