User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/09/15

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Dialysis Test

  1. Prepare 100 mL Glycine Buffer with pH = 3.5
    1. 0.6 mL 0.95 M HCl + 0.37475 g glycine --> pH = 3.27 --> discarded
    2. 0.4 mL 0.95 M HCl + 0.37513 g glycine --> PH = 3.44 --> used with lysozyme
    3. 0.4 mL 0.95 M HCl + 0.37576 g glycine --> pH = 3.45 --> used with 30:1 AuNP - lysozyme
  2. Prepare 100 mL saline solution
    • 0.9 wt-% = 150 mM NaCl
    • 0.90066 g NaCl, 100.00xxx g total --> pH = 5.14
  3. Prepare 90 mL 50 mM NaCl (by dilution)
  4. Prepare 25 mL 1.00 g/L lysozyme
  5. Prepare 4 mL 0.253 mM HAuCl4 (10x dilution of stock) --> pH = 3.41
  6. Save portion of 50 mM dialysis solutions & lysozyme solution
  7. Transfer ~80 mL stock solution to a 400 mL plastic beaker
  8. Cut 3500 MWCO dialysis tubing and wet with DI water
  9. Clamp one end, add 10 mL of 1 g/L lysozyme, clamp other end & place in 50 mM NaCl
  10. Repeat, but place in 50 mM glycine buffer (in beaker)
  11. Repeat, but use ~9 mL 30:1 AuNP - lysozyme colloid (inside) & 50 mM glycine buffer (in beaker)
    • one side loosened, xfer solution to test tube, reclamp and transfer back to tubing
    • only about 6 - 7 mL left after xfers
    • rinsed exterior with DI water prior to soak in buffer



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