All lanes : Anti-LMAN1 antibody [OTI1B8] (ab118407) at 1/200 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : LMAN1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 58 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab118407 observed at 55 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

ab118407 was shown to react with LMAN1 in wild-type HEK-293T cells in western blot with loss of signal observed in LMAN1 knockout cell line ab266248 (LMAN1 knockout cell lysate ab257505). Wild-type and LMAN1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab118407 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 200 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-LMAN1 antibody [EPR6980] (ab126720) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : LMAN1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 58 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab126720 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab126720 Anti-LMAN1 antibody [EPR6980] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab126720 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°CC 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-LMAN1 antibody [EPR6979] (ab125006) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : LMAN1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 58 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab125006 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab125006 Anti-LMAN1 antibody [EPR6979] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab125006 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°CC 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 1

Representative images of LMAN1 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.