Knight:NuPAGE electrophoresis/Fast staining

Overview

A fast protocol for visualizing bands on a polyacrylamide gel via coomassie-like staining.

Materials

• Deionized water
  • Invitrogen SimplyBlue SafeStain
• Microwave
• Plastic tray
• Orbital shaker
• Kimwipes

Procedure

1. Add 100mL deionized water in staining tray.
2. Microwave staining tray loosely covered for 1 min on high.
3. Shake staining tray for 1 min on orbital shaker at room temperature.
4. Discard water.
5. Repeat steps 1-4 two more times.
6. Add 20mL SimplyBlue SafeStain.
   • Need 100mL for staining tray.
7. Microwave staining tray loosely covered for 45 secs on high.
   • 1 min seemed a bit too long.
8. Shake staining tray for 5 mins on orbital shaker at room temperature.
9. Discard SimplyBlue SafeStain.
10. Add 100mL deionized water.
11. Add a kimwipe.
12. Shake staining tray for 10 mins on orbital shaker at room temperature.
13. Add 20mL 20% NaCl for at least 5 mins.
    • Only do this step if you aren't planning on drying the gel! Otherwise, when the gel dries, you will see a white salt precipitate.
14. Gel can be stored in salt solution for several weeks.

This protocol seems to work pretty well but is not as sensitive as the Knight:NuPAGE electrophoresis/Slow staining approach.