Knight:NuPAGE electrophoresis/Slow staining

Overview

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

Materials

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

Procedure

1. Add 100mL deionized water in staining tray.
2. Shake staining tray for 5 min on orbital shaker at room temperature.
3. Discard water.
4. Repeat steps 1-3 two more times.
5. Add 20mL SimplyBlue SafeStain.
6. Shake staining tray for 1 hr on orbital shaker at room temperature.
   • Overnight is better for improved sensitivity.
7. Add 100mL deionized water.
8. Shake staining tray for 1 hr or more on orbital shaker at room temperature.
   • If the gel has been stained overnight, let it destain for a few hours.

This protocol worked less well for me. However, I learned later that the typical trick people use is to include a kimwipe in the destain step to help soak up the stain. This will likely improve the signal to noise on the gel.