User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/07/24

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Procedure

  1. The elutes were placed in the centrifuge again for further concentration (until volume = ~10mL)
  2. The final volume was 9mL
  3. A 1/100th dilution of this solution was prepared and a UV-Vis spectra was taken of it
  4. Using Beer's Law the concentration of protein was calculated
    1. Molar Absorptivity at 415nm: 128,000
    2. Absorbance at 415nm : 0.036
    3. The concentration of protein (not in diluted sample) was calculated to be 28.125uM
    4. This corresponds to 2.53125 * 10^-7 moles of Hemoglobin
  5. Calculate the amount of ruthenium need for a 10:1 ratio
    1. Amount of ruthenium needed : 2.53125 *10^-6 moles
    2. Mass of ruthenium needed : ~1.898mg
  6. Calculate the amount of dithiothreitol needed to make 300uM in Tris
    1. Amount of DTT needed : 0.00003 moles
    2. Mass of DTT needed : 46.2759mg