Lane 1 : mGluR5 antibody at 1/600 dilution Lanes 2 - 4 : Anti-Metabotropic Glutamate Receptor 5 (phospho S996) antibody ( ab16653 ) at 3 µg/ml Lane 1 : mouse hippocampal lysate Lane 2 : mouse hippocampal lysate Lane 3 : mouse hippocampal lysate with Rat Metabotropic Glutamate Receptor 5 (phospho S996) peptide ( ab17573 ) at 3 µg/ml Lane 4 : mouse hippocampal lysate with Rat Metabotropic Glutamate Receptor 5 peptide (ab17575) at 3 µg/ml Lysates/proteins at 10 µg per lane. Performed under reducing conditions. Predicted band size : 130 kDa Observed band size : ~130 kDa ( why is the actual band size different from the predicted? ) Additional bands at : >200 kDa. We are unsure as to the identity of these extra bands. Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom