Anti-CDC42 antibody [M152] (ab41429)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: CDC42 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab41429 observed at 20 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab41429 was shown to specifically react with CDC42 in wild-type HAP1 cells. No band was observed when CDC42 knockout samples were examined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab41429 and ab181602 (loading control to GAPDH) were diluted at 1/125 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

ab41429 staining CDC42 in SH-SY5Y cells by Immunocytochemistry/ Immunofluorescence. SH-SY5Y cells were differentiated 6 days with 10 µM retinoic acid and fixed with 4% paraformaldehyde in PBS. After 5 minutes incubation with ice-cold methanol the cells were washed twice with PBS and blocked with 10% normal donkey serum for 30 minutes. Incubation with primary antibody was performed over night at 4°C. After three-times washing with PBS, 1 hour incubation with the labeled Cy3 secondary antibody at 37°C the cells were mounted with DABCO/Mowiol.

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PC-12 (Rat adrenal gland pheochromocytoma cell line) cells grown for 4 days on poly-D-lysine-coated plates in the rpesence (200 ng/ml) or absence (control) of Nerve Growth Factor (NGF) labeling CDC42 with ab41429 in ICC/IF. Primary antibody was used at 1/50 dilution followed by a donkey anti-mouse secondaty antibody (Cy2).