Human CLOCK (KAT13D) knockout HeLa cell lysate

• WB

Lab

Western blot - Human CLOCK (KAT13D) knockout HeLa cell lysate (AB258365)

Lane 1 : Wild-type HeLa cell lysate 20 μg

Lane 2 : CLOCK knockout HeLa cell lysate 20 μg

Lane 3 : A431 cell lysate 20 μg

Lane 4 : U-251 MG cell lysate 20 μg

False colour image of Western blot : Anti-KAT13D / CLOCK antibody staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab93804 was shown to bind specifically to KAT13D / CLOCK. A band was observed at 100 kDa in wild-type HeLa cell lysates with no signal observed at this size in CLOCK knockout cell line ab266054 (knockout cell lysate ab258365). The band observed in the knockout lysate lane below 100 kDa is likely to represent a truncated form of KAT13D / CLOCK. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CLOCK knockout HeLa 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-KAT13D / CLOCK antibody (<a href='/en-us/products/primary-antibodies/kat13d-clock-antibody-ab93804'>ab93804</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CLOCK knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CLOCK (KAT13D) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-clock-kat13d-knockout-hela-cell-line-ab266054'>ab266054</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

U-251 MG cell lysate at 20 µg

Predicted band size: 95 kDa

Observed band size: 100 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human CLOCK (KAT13D) knockout HeLa cell lysate (AB258365)

Homozygous : 1 bp insertion in exon11