All lanes : Anti-beta Dystroglycan antibody [EP2200Y] (ab62373) at 1/500 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : DAG1 knockout HEK293T cell lysate
Lane 3 : Mouse heart tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 97 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab62373 observed at 43 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab62373 Anti-beta Dystroglycan antibody [EP2200Y] was shown to specifically react with beta Dystroglycan in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266263 (knockout cell lysate ab257192) was used. Wild-type and beta Dystroglycan knockout samples were subjected to SDS-PAGE. ab62373 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.

 

All lanes : Anti-beta Dystroglycan antibody (ab43125) at 1/500 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : DAG1 knockout HEK293T cell lysate
Lane 3 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 97 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab43125 observed at 43 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab43125 Anti-beta Dystroglycan antibody was shown to specifically react with beta Dystroglycan in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266263 (knockout cell lysate ab257192) was used. Wild-type and beta Dystroglycan knockout samples were subjected to SDS-PAGE. ab43125 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: 1 bp insertion in exon 2

Representative images of DAG1 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.