All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : AXL knockout HeLa cell lysate
Lane 3 : NCI-H1299 cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 98 kDa
Observed band size: 80140 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab215205 observed at 80,140 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab215205 Anti-Axl antibody [EPR21107] was shown to specifically react with Axl in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265392 (knockout cell lysate ab257152) was used. Wild-type and Axl knockout samples were subjected to SDS-PAGE. ab215205 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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 10000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 1 bp insertion in exon 4.

Representative images of AXL knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa 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.