人BCKDHA knockout HEK-293T cell裂解物

• WB

Lab

Western blot - Human BCKDHA knockout HEK-293T cell lysate (AB258324)

Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : BCKDHA knockout HEK-293T cell lysate 20 μg
Lane 3 : Human Heart cell lysate 20 μg
Lane 4 : Daudi cell lysate 20 μg
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 (<a href='/products/primary-antibodies/bckdha-antibody-ab126173'>ab126173</a>) 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='/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

false

• Sanger seq

Unknown

Sanger Sequencing - Human BCKDHA knockout HEK-293T cell lysate (AB258324)

Homozygous : Insertion of the selection cassette in exon1