All lanes : Anti-ITPA antibody [EPR8779] (ab150420) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ITPA knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 21 kDa
Observed band size: 21 kDa

Lanes 1-3: Merged signal (red and green). Green - ab150420 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab150420 Anti-ITPA antibody [EPR8779]  was shown to specifically react with ITPA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266679 (knockout cell lysate ab258477) was used. Wild-type and ITPA knockout samples were subjected to SDS-PAGE. ab150420 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-ITPA antibody [EPR8780] (ab134937) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ITPA knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 21 kDa
Observed band size: 21 kDa

Lanes 1-3: Merged signal (red and green). Green - ab134937 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab134937 Anti-ITPA antibody [EPR8780]  was shown to specifically react with ITPA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266679 (knockout cell lysate ab258477) was used. Wild-type and ITPA knockout samples were subjected to SDS-PAGE. ab134937 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 1

Representative images of ITPA 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 an EVOS M5000 microscope.