1. Cell lysis: breaks down cells to isolate proteins.
Mercaptoehtanol treatment breaks disulfide
linkages to denature protein and have it unfolded.
2. Protein separation on a gel:sodium dodecyl
sulfate added to give uniform negative charge to
proteins and current puls protein down get to
positive chargeProteins are separated based on
size: big at the top, small at the botton
Protein Detection: use antibodies to detect proteins at specific molecular weight
- protein size is measured in kilodaltons
→ Pros: can test multiple proteins, proteins easier to detect by antibody as unfolded
→ Cons: No spatial info