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