User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/10/13

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SDS-PAGE Sample Buffer

  • 2X recipe: 0.125 M Tris (pH adjusted to 6.8), 4% SDS, 20% glycerol, 0.01% bromophenol blue, & 5% 2-ME or 0.15 M DTT
  • For 10 mL, no disulfide bond breaker
    • 0.151 g Tris
    • 0.40 g SDS
    • 2 g glycerol
    • 0.001 g bromophenol blue
    • 7.5 mL H2O - mL HCl required to make pH 6.8
  • Actual prepared sample
    • 7 mL H2O
    • 0.151 g Tris
    • ~2 mL 0.5 N HCl (ran out of 1 M HCl) -- pH 6.7
    • 0.40 g SDS
    • 0.001 g bromophenol blue (no DMSO available, but dissolved quickly this time)
    • 2 g glycerol (added last)
    • stored in 15 mL falcon tube (~11 mL)


Unknown Protein Samples

  1. Soy protein
    • 1 mg in 1.5 mL H2O (centrifuge vial)
    • did not completely dissolve, even with 5 min vortex
    • after settling, supernatant stored
  2. BSA
    • 0.1 - 0.2 mg in 1 mL H2O (centrifuge vial)
    • quickly dissolved - clear, colorless solution
  3. Casein
    • 2.6 mg in 10 mL H2O (falcon tube)
    • did not dissolve after 5 min vortex -- left in vial
  4. Horseradish Peroxidase (HRP)
    • 0.4 mg in 3 mL H2O (falcon tube)
    • quickly dissolved - clear, colorless solution