All lanes : Anti-NR2F2 antibody [EPR18443] (ab211777) at 1/1000 dilution

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : NR2F2 knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 46 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab211777 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab211777 was shown to react with NR2F2 in wild-type HCT116 cells in western blot. The band observed in knockout cell line ab266888 (knockout cell lysate ab257186) lane below 45kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NR2F2 knockout HCT117 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab211777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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: 1 bp insertion in exon2