All lanes : Anti-FOXP3 antibody ( ab15590 ) at 1 µg/ml Lane 1 : HEK293 overexpressing mouse FOXP3 Lane 2 : HEK293 overexpressing human FOXP3 Lane 3 : HEK293 overexpressing mouse FOXP3 with Mouse FOXP3 peptide (ab15736) at 1 µg/ml Lane 4 : HEK293 overexpressing human FOXP3 with Mouse FOXP3 peptide (ab15736) at 1 µg/ml Lysates/proteins at 20 µg per lane. Secondary Goat anti-Rabbit HRP at 1/5000 dilution Performed under reducing conditions. Predicted band size : 50 kDa Observed band size : 52 kDa ( why is the actual band size different from the predicted? ) Exposure time : 3 minutes