User:Madeleine Y. Bee/Notebook/CHEM-571 2013F/2013/09/18

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September 18, 2013


Follow this procedure to perform a redox titration for Horseradish peroxidase and determine it's standard reduction potential.


The redox reaction will be monitored with a spectroelectrochemical cell, and UV-vis measurements will be taken to observe the resulting redox state. DTT will be added in 1uL increments and the UVvis and circuit potential data will be recorded, to eventually plot %oxidized or %reduced versus voltage. <br.>

  • Buffer: 50mM TRIS/50mM NaCl
  • Sodium dithionite: 1.46M
  • Horseradish peroxidase: 33uM
  • Redox mediators (to stabilize the solution potential)dye solution:
    • Diluted 1000x
    1. Duroquinone: 33.3mg in 10mL --> 20.3mM
    2. 2-hydroxy-1,4-naphthoquinone 18.0mg in 10mL --> 10.3mM
    3. anthraquinone-2-sulfonate 30.8mg --> 9.93mM
    4. benzyl viologen 8.7mg --> 2.1mM
    5. phenylsafranine 2.6mg --> 0.81mM
    6. indigo-carmine 4.6mg --> 0.99mM