All lanes : Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : FLOT2 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : A549 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: 47 kDa
Observed band size: 47 kDa

Lanes 1-4: Merged signal (red and green). Green - ab181988 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab181988 Anti-Flotillin 2/ESA antibody [EPR14128(B)] was shown to specifically react with Flotillin 2/ESA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266847 (knockout cell lysate ab257442) was used. Wild-type and Flotillin 2/ESA knockout samples were subjected to SDS-PAGE. ab181988 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.

Homozygous: 5 bp deletion in exon4