Lidstrom: SDS-PAGE

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Gel Prep

• Clean cover plate and thicker spacer plate 75 mM
  • Soap and Water
  • Ethanol
  • DI water
• Dry plates
• Setup one spacer plate and one cover plate in each gel holder
  • The cover plate goes on the side of the spacer plate with the spacers in order to create a small gap between the plates
• Put the gel holder into the casting stand

Pour the Resolving Gel

• Mix components of the amounts in the Gel Mix link. Mix in the order listed.
• Gel Mix Recipe
  • Don't add APS/TEMED until ready to pour
• Use pipette to put gel mix into the gap between the plates
• Carefully layer 50%EtOH 50% ddH2O on top of the gel to prevent the top of the gel from drying out
• Let dry for an hour

Pour the Stacking Gel

• Mix components of the amounts in the Gel Mix link. Mix in the order listed.
• Gel Mix Recipe
  • Don't add APS/TEMED until ready to pour
• Use pipette to put gel mix into the gap between the plates
• Insert the comb being careful not to trap any bubbles
• Let dry for an hour

Sample Prep

• 10-20 ug protein
• Load 12-15 uL, absolute max 20 uL for big comb
• Denature protein by mixing with sample buffer, BME and boiling

Other Resources

• Background
• Instructions for PageRuler protein standard