Knight:NuPAGE electrophoresis/Hybrid staining
A fast protocol for visualizing bands on a polyacrylamide gel via coomassie-like staining.
- Deionized water
- Invitrogen SimplyBlue SafeStain
- Plastic tray
- Use lid of a 1000μL pipette tip box.
- Orbital shaker
- Add 100mL deionized water in staining tray.
- Microwave staining tray loosely covered for 30 secs on high.
- Shake staining tray for 1 min on orbital shaker at room temperature.
- Discard water.
- Repeat steps 1-4 two more times.
- Add 20mL SimplyBlue SafeStain (enough to just cover the gel).
- Microwave staining tray loosely covered for 20 secs on high.
- Shake staining tray for 5 mins on orbital shaker at room temperature.
- You should now be able to see most of the bands on the gel.
- Place staining tray with gel inside a ziploc bag to limit evaporation.
- Incubate overnight on orbital shaker at room temperature.
- Overnight incubation will yield better sensitivity due to darker staining.
- Discard SimplyBlue SafeStain.
- Add 100mL deionized water.
- Add 2 kimwipes.
- Shake staining tray for 2-3 hours on orbital shaker at room temperature.
- Take gel picture.
- 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.
- Gel can be stored in salt solution for several weeks.