SDS PAGE and staining buffers

10X SDS-PAGE running buffer

• 30.3 g Tris base
• 144.0 g Glycine
• 10.0 g SDS

This makes 1 L – scale up for more volume. No need to adjust pH.

Coomassie Staining

from: Studier, F., Protein Expression and Purification, 1995

• Stain solution 1: 50% EtOH, 10% acetic acid
• Stain solution 2: 5% ethanol, 7.5% acetic acid
• Coomassie stock solution: 0.25% Coomassie brilliant blue R250 in 95% ethanol

Protocol:

1. Place gel in 50 ml of Stain solution 1, microwave for 25 sec.
2. Rock for 10 min.
3. Place gel in 50 ml of Stain solution 2, add 250 µl Coomassie, and microwave for 35 sec.
4. Rock for 1 hour to overnight.