User:Moira M. Esson/Notebook/CHEM-571/2013/11/05

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Objectives

  • Analyze protein-AuNPs synthesized on 10/22/2013 and 10/23/2013 using Atomic Absorption spectroscopy


AA spectroscopy

General Protocol:

  1. Transfer ~4mL of sample from test tubes into a 10mL falcon tube.
  2. Centrifuge ~4mL protein-AuNP solutions for 30 minutes at 4000rpm. Be sure that the centrifuge is balanced.
  3. Prepare 5 standard Au solutions (10μg/mL, 20μg/mL, 30μg/mL, 40μg/mL, 50μg/mL) following protocol described on 09/10/2013.
  4. Run samples on AA.
  5. Prepare calibration curve and prepare a graph showing the concentration of Au vs Au:Protein ratio


Table 1. 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)
100.19.9
200.29.8
300.39.7
400.49.6
500.59.5


Figure 1. AA Gold Standard Calibration Curve
Image:CalibrationcurveAAmyohemozem.png
Figure 2. Hemoglobin Absorbance Curve
Image:Concentration vs Absorbance Hemoglobin.jpg
Figure 3. Myoglobin Absorbance Curve
Image:Concentration vs Absorbance Myoglobin.jpg

Notes

  • Lysozyme solutions prepared on 10/30/2013 all produced fibers and appear colorless except for the 30:1 ratio(appears furthest to the right). This solution is mildly colored.
  • All myoglobin solutions prepared on 10/30/2013 produced fibers and appear colorless.




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