All lanes : Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (ab109252) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : S100A13 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 11 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab109252 observed at 13 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab109252 was shown to react with S100 Calcium Binding Protein A13/S100A13 in wild-type HEK-293T cells in Western blot with loss of signal observed in S100A13 knockout cell line ab266065 (S100A13 knockout cell lysate ab258183). Wild-type HEK-293T and S100A13 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab109252 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 h at room temperature before imaging.

Homozygous: Insertion of the selection cassette in exon 4