Knight:NuPAGE electrophoresis/Fast staining

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(Materials)
(Materials)
Line 7: Line 7:
*Microwave
*Microwave
*Plastic tray
*Plastic tray
 +
**''Use lid of a 1000μL pipette tip box.''
*Orbital shaker
*Orbital shaker
*Kimwipes
*Kimwipes

Revision as of 19:36, 16 January 2007

Overview

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

Materials

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.

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