I'm a Biomedical Science honours student. I have a western blot dilemma so I would love any help I can get. I study tetraspanin proteins, in particular CD9. The CD9 antibody I have requires the protein to be non-reduced and the nitrocellulose blots are fine. However, I then bock and stain with either total actin or beta-tubulin III and instead of a single band around 50, i get 2 or three bands around 50!! Does anyone have any ideas as to why this may be occurring? I thought maybe its picking up different isoforms? Therefore it seems i cannot use these as loading controls??