Knight:NuPAGE electrophoresis/Fast staining
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Revision as of 19:36, 16 January 2007
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 1 min 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.
- Need 100mL for staining tray.
- Microwave staining tray loosely covered for 45 secs on high.
- 1 min seemed a bit too long.
- Shake staining tray for 5 mins on orbital shaker at room temperature.
- Discard SimplyBlue SafeStain.
- Add 100mL deionized water.
- Add a kimwipe.
- Shake staining tray for 10 mins on orbital shaker at room temperature.
- 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.
This protocol seems to work pretty well but is not as sensitive as the Knight:NuPAGE electrophoresis/Slow staining approach.