All lanes : Anti-COMT antibody [EPR6490] (ab126618) at 1/1000 dilution

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

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 30 kDa
Observed band size: 24-28 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab126618 observed at 24-28 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab126618 Anti-COMT antibody [EPR6490] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab126618 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 20000 dilution for 1 hour at room temperature before imaging.

 

All lanes : Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

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

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 30 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab124813 Anti-COMT antibody [EPR6491(B)] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab124813 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 20000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 1 bp insertion in exon 3

Representative images of COMT 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.