User:Paul Rothenberg/Notebook/Pauls Notebook/2014/03/26

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UV-VIS of Apoprotein

  • UV-VIS spectroscopy was run on the protein to determine if the extraction of the heme groups was successful. If so, there should be a peak at 280 nm. If there is heme left, there would be a peak at 400 nm.
  • The spectrum revealed a large peak at 280 nm (roughly 0.7) and a very slight peak at 400 (0.06-0.07).

Reconstitution of Apoprotein

  • In order to reconstitute the apoprotein with the Mn PPIX, the ratio used in other syntheses was 2 mg PPIX per 10 mg apoprotein product (assuming full yield); this is still an excess of PPIX. For this reaction, 200 mg of myoglobin was used, so the stoichiometric amount of PPIX is 40 mg. Of which, we have 500 mg.
  • Next, we need 2 mL of 100mM Phosphate buffer at pH 7.0, and 0.1 N (0.1 M) NaOH.


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