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Single Percentage Polyacrylamide Gels

Recipe for casting common single percentage protein polyacrylamide gels.

Separation Gel (10mL) 7% 10% 12.5% 15% Stacking Gel (4mL)
H2O 5.1 mL 4.1 mL 3.3 mL 2.5 mL 2.1 mL
Acrylamide/bis (30%) 2.3 mL 3.3 mL 4.1 mL 5.0 mL 0.8 mL
Tris-HCL (1.5M, pH 8.8) 2.5 mL 2.5 mL 2.5 mL 2.5 mL 1 mL
10% SDS 100 µL 100 µL 100 µL 100 µL 40 µL
TEMED 10 µL 10 µL 10 µL 10 µL 40 µL
10% APS 32 µL 32 µL 32 µL 32 µL 4 µL
  1. Prepare separation and stacking gels without TEMED and APS.
  2. Add TEMED and APS to separation gels; pour into molds stopping 2cm from the top.
  3. Add isopropanol to the tops of the gels and wait 20-30min for the gels to solidify.
  4. Pour out isopropanol & blot tops of the gels to ensure the isopropanol is fully removed.
  5. Add TEMED and APS to stacking gels; pour on top of solidified separation gel layers and add combs.
  6. Use immediately or wrap in wet paper towels & store at 4°C up to 2 weeks.

10% Ammonium persulphate (APS) solution

  • 1 g APS / 10 mL H2O
  • Store at 4°C up to 6 mo.

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