All lanes : Anti-CD130 (gp130) antibody [EPR21732] (ab217671) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : IL6ST knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 104 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab217671 observed at 130 kDa. Red - loading control ab7291 observed at 50 kDa.

 ab217671 Anti-CD130 (gp130) antibody [EPR21732] was shown to specifically react with CD130 (gp130) in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266938 (knockout cell lysate ab257207) was used. Wild-type and CD130 (gp130) knockout samples were subjected to SDS-PAGE. ab217671 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 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.

 

 

Allele-1: 4 bp deletion in exon4

 

Allele-2: 1 bp insertion in exon 4.

 

Representative images of IL6ST knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 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.