User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/31

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Separate Protein and Lysozyme UV-Vis

Goals

  • Separate the proteins from the cells
  • Run UV-Vis on the AuNP/Lysozyme solutions

Procedure

  • The same procedure from 2012/09/25 was followed in order to separate out the proteins from the cell organelles.
  • For information regarding the AuNP lysozyme UV-Vis spectras please see Michael's Notebook.

Data

UV-Vis runs of AuNP in Lysozyme

Redone samples in Quartz Cuvets

Conclusions

  • The separation process ran smoothly except for during the vacuum filtration. There seemed to be a defect in the clamp being used which caused some of the supernatant collected to leak out. A glass pipette was used to recollect as much as possible and placed back through the filter. This defect may be the cause for any loss in ADA collected when conducting the FPLC.
  • Samples 134-140 showed a variation in the baseline and as a result, in order to determine whether this was due to lysozyme or a variation int he cuvettes, these four trials were repeated using the same quartz cuvet. A blank of water was run using the same cuvette and subtracted from each spectra manually.