User:Beatriz Gimenez De C./Notebook/571/2014/11/04

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Tasks

  • Analysis of data of dialysis set on 9/29/2014, 0.6 g/L Lysozyme vs HCl using MWCO 20000:
    • Bradford and UV-Vis analysis of the 5 Lysozyme containing solutions.
      • used 100 μL of each solution for both experiments
      • Bradford reagent diluted 1:3 with 50mM Tris/50mM NaCl added to Polystyrene cuvette with 100 μL solution + 700 μL Tris/NaCl buffer

Image:11-04-2014 Lysozyme vs HCl Bradford Assay.png

Image:11-04-2014 Lysozyme vs HCl Bradford Assay 2.png

      • UV-Vis 100 μL solutions were diluted100 times and a quartz cuvette was used

Image:11-04-2014 Lysozyme vs HCl UV-Vis.png

    • pH data
Concentration of HCl (μM) pH
75 (1) 7.142
250 (3) 5.707
500 (4) 4.944
5000 (5) 2.996
lysozyme sample pH
Lys 1 7.116
Lys 2 7.423
Lys 3 6.158
Lys 4 4.921
Lys 5 3.025
  • Dialysis preparation:
    • Prepare CaCl2 solutions
      • Prepare stock solution of 45 mM CaCl2 (0.2502 g CaCl2 in 50 mL volumetric flask + HPLC water).
      • Dilute solution to obtain 35, 25, 15 and 5 mM CaCl2 mM solutions.
    • Dialysis setting: 30:1 colloid solution vs CaCl2 using MWCO 3500 dialysis (1 day on low speed steerer).
      • 5 mM CaCl2 vs 30:1 Colloid
      • 15 mM CaCl2 vs 30:1 Colloid
      • 25 mM CaCl2 vs 30:1 Colloid
      • 35 mM CaCl2 vs 30:1 Colloid
      • 45 mM CaCl2 vs 30:1 Colloid



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