All lanes : Anti-RhoGDI antibody [EPR3773] (ab133248) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ARHGDIA knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HeLa 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: 23 kDa
Observed band size: 23 kDa

Lanes 1-4: Merged signal (red and green). Green - ab133248 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab133248 Anti-RhoGDI antibody [EPR3773] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab133248 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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.

Allele-1: 1 bp insertion in exon 2

 

All lanes : Anti-RhoGDI antibody [EPR3772] (ab108977) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ARHGDIA knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HeLa 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: 23 kDa
Observed band size: 23 kDa

Lanes 1-4: Merged signal (red and green). Green - ab108977 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab108977 Anti-RhoGDI antibody [EPR3772] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab108977 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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.

Allele-2: Insertion of the selection cassette in exon 2.