All lanes : Anti-HEF1 antibody ( ab37161 ) at 1/250 dilution Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate ( ab7899 ) Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HEF1 peptide (ab39117) at 1 µg/ml Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate ( ab7899 ) with Human HEF1 peptide (ab39117) 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 : 110 kDa Observed band size : 120 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.