User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/07/14

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  1. Our protein solution was collected from the cold room and transferred into a 50mL falcon tube labeled Purified Hb pH7, 7/14/14
  2. We will run a gel with the solution and past solution to check purity
  3. The gel will be set up in the following order
    1. Myoglobin and BSA ladder
    2. Empty
    3. 1/10 dilution of Protein in pH8.3 tris (solution that was injected into Q-Sepharose column)
    4. Elute during injection of Protein in pH8.3 tris into Q-Sepharose column
    5. Elute during cleaning Q-Sepharose column containing Protein in pH8.3 tris with 1M NaCl
    6. Elute from SP-Sepharose column in phosphate buffer pH7.2 during injection (from fractions collected from Q-Sepharose column)
    7. Empty
    8. 1/10 dilution of Ammonium sulfate treatment pellet dissolved in 50mM Tris
    9. 1/10 dilution of purified protein from dialysis over the weekend
    10. 1/100 dilution of purified protein from dialysis over the weekend
    11. 1/10 dilution of centrifuged purified protein from dialysis over the weekend
    12. Myoglobin and BSA ladder
  4. The gel was run at 200V for 34 minutes in 1X Running Buffer

Stock Solutions

  1. Protein ladder

-1mg BSA and 1mg Myoglobin in 1mL water -Solution was diluted to 1/10 to be used in gel

  1. 1X Running Buffer

-14.4g Glycine, 1g SDS, 3g Tris in 100mL H2O -Solution was diluted to 1L with water

  1. Fixative Solution

-40 Methanol, 10% Acetic Acid, 50% Water

  1. Stain Solution

-90% water, 10$ Acetic Acid and .0025% Commassie Brilliant Blue

  1. Destain Solution

-90% Water and 10% Acetic Acid



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