All lanes : Anti-Cortactin antibody [EP1922Y] (ab81208) at 1/50000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CTNN knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab81208 observed at 70 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab81208 Anti-Cortactin antibody [EP1922Y] was shown to specifically react with Cortactin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266819 (knockout cell lysate ab257147) was used. Wild-type and Cortactin knockout samples were subjected to SDS-PAGE. ab81208 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 50000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 2 bp deletion in exon 3

Representative images of CTTN knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.