All lanes : Anti-SGK1 antibody ( ab43606 ) at 1 µg/ml Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human SGK1 peptide (ab43605) at 1 µg/ml Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human SGK1 peptide (ab43605) at 1 µg/ml Lysates/proteins at 10 µg per lane. Secondary IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution Performed under reducing conditions. Predicted band size : 49 kDa Observed band size : 68 kDa ( why is the actual band size different from the predicted? ) SGK1 contains Glu/Lys-rich regions and a number of phosphorylated Ser/Thr sites (SwissProt data) which may explain why the observed band is running higher than the predicted molecular weight.