All lanes : Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/1000 dilution

Lane 1 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
Lane 2 : T-47D (Human ductal breast epithelial tumor cell line) cell lysate
Lane 3 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 4 : MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Observed band size: 24 kDa why is the actual band size different from the predicted?

Lanes 1- 4: Merged signal (red and green). Green - ab45167 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab45167 was shown to react with MUC1 in Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab255412 (knockout cell lysate ab263764) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) 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. ab45167 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 exon 1.