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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 |
- Prepare separation and stacking gels without TEMED and APS.
- Add TEMED and APS to separation gels; pour into molds stopping 2cm from the top.
- Add isopropanol to the tops of the gels and wait 20-30min for the gels to solidify.
- Pour out isopropanol & blot tops of the gels to ensure the isopropanol is fully removed.
- Add TEMED and APS to stacking gels; pour on top of solidified separation gel layers and add combs.
- 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.