Team:Aachen/Wetlab/Protocol33

SDS page gel

Preparation of the separation-gel (T=12%), pH 8.8:


  1. mix:

    • 2.115 ml ddH2O
    • 3.75 ml Tris-HCl-Stock (1M, pH 8.8)
    • 4 mL AA-Stock (30%)
    • 100uL SDS (10%) (mix well afterwards)
    • 10 uL TAMED
    • 30 uL APS (20%)

  2. mix all components, without the APS
  3. add APS and pipette 4 ml between the glass panes
  4. overlay the gel with isopropanol and wait for 30 min for the gel to polymerize

Preparation of the stacking-gel (T=4%), pH 6.8:


  1. mix:

    • 3.645 ml ddH2O
    • 625 ul Tris-HCl-Stock (1M, pH 6.8)
    • 830 ul AA-Stock (30%)
    • 50 ul SDS (10%) (mix well afterwards)
    • 5 ul TAMED
    • 15 ul APS (20%)

  2. mix all components, without the APS
  3. remove the isopropanol with a filter paper
  4. add APS and pipette it on top of the separation-gel
  5. put the comb into the liquid gel
  6. wait for 30 min until the gel is polimerized
  7. if not used immediately, wrap into wet cloth and put into the fridge for storage (optional)