All lanes : Anti-ETFA antibody [2B11AE8] (ab110316) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : ETFA knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate

Lysates/proteins at 40 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

Predicted band size: 35 kDa
Observed band size: 35 kDa

Lanes 1-2: Merged signal (red and green). Green - ab110316 observed at 35 kDa. Red - loading control ab52901 observed at  kDa.

 ab110316 Anti-ETFA antibody [2B11AE8]  was shown to specifically react with ETFA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266513 (knockout cell lysate ab257943) was used. Wild-type and ETFA knockout samples were subjected to SDS-PAGE. ab110316 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) 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^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Homozygous: Insertion of the selection cassette in exon 1