All lanes : Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (ab125001) at 1/1000 dilution

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : RXRA knockout HCT116 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MCF7 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 51 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab125001 observed at 53 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab125001 was shown to react with Anti-Retinoid X Receptor alpha in wild-type HCT 116 cells in western blot with loss of signal observed in RXRA knockout cell line ab273708 (RXRA knockout cell lysate ab275245). Wild-type and RXRA knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab125001 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Allele-1: 1 bp insertion in exon 1.