Lane 1: Wild-type HeLa cell lysate (20µg)

Lane 2: PAK1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab223849 observed at 65 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab223849 Anti-PAK1 antibody [EPR20048] was shown to specifically react with PAK1 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264889 (knockout cell lysate ab257572) lane below 65kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PAK1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab223849 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 1000 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: 5 bp deletion in exon 2

 

Allele-2: 2 bp deletion in exon 2

 

Allele-3: Insertion of the selection cassette in exon 2