Knight:NuPAGE electrophoresis/Slow staining: Difference between revisions
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*[http://www.invitrogen.com/content.cfm?pageid=9869 Invitrogen SimplyBlue SafeStain] | *[http://www.invitrogen.com/content.cfm?pageid=9869 Invitrogen SimplyBlue SafeStain] | ||
*Plastic tray | *Plastic tray | ||
**''Use lid of a 1000μL pipette tip box.'' | |||
*Orbital shaker | *Orbital shaker | ||
*Kimwipes | *Kimwipes | ||
Revision as of 16:36, 16 January 2007
Overview
A slow protocol for visualizing bands on a polyacrylamide gel via coomassie-like staining.
Materials
- Deionized water
- Invitrogen SimplyBlue SafeStain
- Plastic tray
- Use lid of a 1000μL pipette tip box.
- Orbital shaker
- Kimwipes
Procedure
- Add 100mL deionized water in staining tray.
- Shake staining tray for 5 min on orbital shaker at room temperature.
- Discard water.
- Repeat steps 1-3 two more times.
- Add 20mL SimplyBlue SafeStain.
- Shake staining tray for 1 hr on orbital shaker at room temperature.
- Overnight is better for improved sensitivity.
- Add 100mL deionized water.
- 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.
