User:Omar Choudary/Notebook/Omar Choudary CHEM-571 AU-2011/2012/2012/03/07

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Objective

  • Terminate tagging reaction
  • Prepare solutions for dialysis

Description

  • BSA control solution (BSA/HCl):

Added 1µL HCl (1M)

pH = 6.59 and the solution turned pink

  • 166 ratio solution:

Added 37µL HCl (1M)

pH = 5.53 and the solution turned light pink

  • 70 ratio solution:

Added 5µL of HCl (1M)

pH = 6.83 and the solution turned light purple

  • Began dialysis in water. Dialysis was performed using regenerated cellulose (Fisherbrand) dialysis tubing, in the dark (covered by foil).
  • Dialysis Water Changes:

1. 11:30 am and 5:20 pm on 3/8

2. 2:30 pm on 3/9

3. 10:15 am on 3/10

4. 10:45 am on 3/11

5. 2 pm on 3/12

6. 10 am on 3/14

Notes

Compared to the previous tagging experiment, to terminate the reaction, HCl was added until the solution turned pink/purple and no Tris buffer would be added to raise the pH at all. Tris buffer will be added after performing the dialysis, just before performing the fluorescence study, and until the solution turns blue again. There will be a significant reduction in the amount of Tris added to the solution compared to the previous tagging experiment. There is the hope that this will yield more accurate results regarding the tagging of the lysine.