All lanes : Anti-Securin antibody [EPR3240] (ab79546) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : PTTG1 knockout HEK293T cell lysate
Lane 3 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 22 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab79546 observed at 28 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab79546 Anti-Securin antibody [EPR3240] was shown to specifically react with Securin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266231 (knockout cell lysate ab257289) was used. Wild-type and Securin knockout samples were subjected to SDS-PAGE. ab79546 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.

 

All lanes : Anti-Securin antibody (ab26273) at 1/500 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : PTTG1 knockout HEK293T cell lysate
Lane 3 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 22 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab26273 observed at 28 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab26273 Anti-Securin antibody was shown to specifically react with Securin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266231 (knockout cell lysate ab257289) was used. Wild-type and Securin knockout samples were subjected to SDS-PAGE. ab26273 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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: 8 bp deletion in exon 1

Representative images of PTTG1 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.