User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/07/22

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Q-Sepharose column

The remaining protein solution was placed in the fridge with 50mM Tris buffer at pH 8.3

  1. The Q-sepharose column was attached to the FPLC
  2. The protein solution was loaded in 50mL at a time
    1. the elute was collected during injection
  3. After each 50mL, a gradient of 0-160mM NaCl was run in Tris pH 8.3
    1. Fractions were collected during the gradient
      1. For the first 50mL, fractions 4-10 were collected
      2. For the second 50mL, fractions .......

Fractions 1-3 and ....

  1. Between each injection, the column was cleaning with 1M NaCL and reequilibrated with 20mM Tris at pH8.3


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