Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: EZR knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab133297 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133297 Anti-Ezrin antibody [EPR803] was shown to specifically react with Ezrin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264912 (knockout cell lysate ab256908) was used. Wild-type and Ezrin 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. ab133297 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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^® 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.

Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: EZR knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab75840 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab75840 Anti-Ezrin antibody [EP924Y] was shown to specifically react with Ezrin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264912 (knockout cell lysate ab256908) was used. Wild-type and Ezrin 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. ab75840 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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^® 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: 2 bp deletion in exon 6

Allele-2: Insertion of the selection cassette in exon 6

Allele-3: Insertion of the selection cassette in exon 6