User:Monika Gasiorek/Notebook/CHEM-571 2014F/2014/09/24

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September 24, 2014

Bradford Analysis of Dialysis

A Bradford analysis was done on the dialysis samples prepared and dialyzed last week.

  • 600 µL of each soak solution were combined with 200 µL of Bradford reagent and 200 µL of a 50 mM Tris/NaCl buffer
    • The soak solutions included:
                              50 mM NaCl stock solution with 3500 g/mol MWCO tubing
                              50 mM Glycine stock solution with 3500 g/mol MWCO tubing
                              Pure water with 3500 g/mol MWCO tubing 
                              50 mM Glycine stock solution with 25k g/mol MWCO tubing
                              50 mM Glycine stock solution with 3500 g/mol MWCO tubing
  • 100 µL of the contents of each dialysis tubing was combined with 200 µL of Bradford reagent and 700 µL of Tris/NaCl buffer
    • The respective dialysis bag contents were analyzed, all of which initially included 4 mL of 1g/L lysozyme stock solution, except for the very last system, in which 4.0 mL of 36.5 [Au]:[Lys] colloid solution was used in the place of lysozyme against the 50 mM glycine soak with 3500 tubing

NOTE: As mentioned previously, the contents of the tube in the fourth system (50 mM glycine solution with 25K g/mol MWCO tubing) were lost during transfer; thus, only four tubing samples are included in the results.

Solution Conc of Lysozyme (µg/mL) Abs
3500 NaCl Soak -3.344660194 0.028
3500 H2O Soak -3.975728155 0.015
3500 Gly Soak 2.529126214 0.149
25k Gly Soak -0.09223301 0.095
3500 Gly Colloid 1.703883495 0.132
3500 NaCl/Lys Bag 17.91747573 0.466
3500 H2O/ Lys Bag 18.54854369 0.479
3500 Gly/Lys Bag 20.05339806 0.51
3500 Gly/Coll Bag 2.82038835 0.155




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