Hi, I have been running western blots on the same cell line but consistently get a smudge in some of the bands. Aside from air bubbles potentially existing between the membrane and the gel during the transfer (which I try to take care of), I don’t know what else could be going wrong. Any suggestions?
Hi Sachin
There could be multiple reasons for the same:
- Protein Overload
- Too much salt in your sample
- Air Bubbles in the Gel
- partially polymerized gels (if your APS or TEMED are old or Temperature too low) Let us know more details and we might be able to help you better
Hi Sachin
It can also be caused by Poor transfer conditions or Antibody related issue. Are you wetting your membrane in buffer adequately? I can help more if you can share images or more details on your protocol?