User:Matt Hartings/Notebook/Photosynthesis/2012/06/26
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ObjectivePurify Hemoglobin extracted and dialyzed yesterday and extract and dialyze WT Hb that Tamra expressed. DescriptionPurifying WT HbThe wt hemoglobin was extracted at very high concentrations (i.e. the extract was a deep red color). I had roughly 70 mL to purify. I ran 10 mL at a time through the column.
The two sets of FPLC runs were separated because I took a break for lunch and didn't want the UV detector running while I was gone. The desired protein fractions were pooled and stored at 4C. After the final elution I cleaned the system.
Extracting Tamra's Hemoglobin ExpressionTamra had expressed hemoglobin, and, instead of using hemin in DMF, she used a hemin stock that I had made. The hemin stock was prepared by:
The hope with doing this was that the expression would work as well, and there wouldn't be lots of hemin and hemin byproducts left after centrifuging, which seem to be causing my Q columns to turn black. Extraction
Note: the extract here was not as red as the extract that I had when adding hemin in DMF at induction. Data
NotesThis area is for any observations or conclusions that you would like to note.
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