User:Eleni N. Kalivas/Notebook/CHEM-571/2013/11/05

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Objective

will be doing AA on their samples from last week (HRP-AuNPs and pepsin-AuNPs) and this week (Myoglobin-AuNPs and HRP-AuNPs).

Protocol

Atomic Absorption

    1. centrifuge (4mL of your) samples (use the large centrifuge upstairs) 30min at 4000rpm.
    2. make standard solutions in the same manner as the last time we performed AA. But, this time make concentrations that go up to 50ug/mL
    3. Take measurements of your standards and samples
    4. Make a graph showing concentration of Au vs Au:protein ratio

Standard Au Solution preparation (from 1000μg/mL AA Au gold standard solution)

Concentration(μg/mL) Volume of stock Au solution(mL) Volume Deionized water added(mL)
10 0.1 9.9
20 0.2 9.8
30 0.3 9.7
40 0.4 9.6
50 0.5 9.5

Data

  • Atomic Absorbance Gold Standard Calibration Curve

CalibrationcurveAAmyohemozem.png

  • Hemoglobin Absorbance Curve

Concentration vs Absorbance Hemoglobin.jpg

  • Myoglobin Absorbance Curve

Concentration vs Absorbance Myoglobin.jpg

Additional Observations

Lysozyme solutions all produced fibers and appear colorless except for the 30:1 ratio, which contained light pigmentation. All myoglobin solutions produced fibers and appear colorless.