User:Beatriz Gimenez De C./Notebook/571/2014/10/15

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Tasks

  • Performed data analysis of dialysed solutions set October 14, analysis described on october 14
  • Khyra performed titrations of the dialyzed KI samples. Procedure noted on Khyra's notebook
  • Run Electrophoresis
    • Warm dialyzed samples from october 14 to about 40 °C using heating block
    • Transfer solutions to a pre-made gel
      • Well 1: Precision Plus Protein All Blue Standards
      • Well 3: 0.12 g/L lysozyme Ink A
      • Well 4: 30:1 Lysozyme colloid
      • Well 5: 0.12 g/L Lysozyme
      • Well 6: 0.6 g/L Lysozyme
    • Run for 40 minutes at 200V
    • Develop/Stain your gel
      • Place gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes
      • Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour
      • Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes (Repeat this step with fresh destain solution 2 more times)
  • Determination of molecular weight
    • By comparison with Precision Plus Protein Standards, the molecular weight of all samples was close to 15 KD, slightly lower, as expected for lysozyme compounds with MW=14,300 KD.
    • There was a very small quantity of unknown A and colloid in the gel. The very faint mark observed in well 3 could even be considered to be bleed-over from the 30:1 colloid in well 4. Therefore, electrophoresis was inclonclusive with respect to the molecular weight of unknown A.

Electrophoresis gel 151014.jpg

Precision Plus Protein.jpeg