All lanes : Anti-MDC1 antibody ( ab41951 ) at 1 µg/ml Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Mouse MDC1 peptide (ab41950) 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 : 185 kDa Observed band size : 238 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 50 kDa. We are unsure as to the identity of these extra bands.