Knight:NuPAGE electrophoresis/Slow staining

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(Materials)
Line 6: Line 6:
*[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 19:36, 16 January 2007

Overview

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

Materials

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.

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