User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/09/28

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FPLC was performed on the protein extracted from cells


  1. The protein from the dialysis tubes were filtered
  2. FPLC was used to purify the protein.

FLCP Directions

  1. System equilibration was performed with 50 mM Tris (pH 9)
  2. Protein was loaded onto the loop
  3. Buffer B ( 50 mM tris + 1 M NaCl) was run through the system. Buffer A (50 mM Tris) was run through the system after Buffer B.
  4. The column was injected with protein.
  5. The computer was set to collect 5 mL/min
  6. The column was washed again with Buffer A
  7. The gradient (50% buffer B in 15 minutes) was run and fractions were collected
  8. The fractions for each peak were collected and later divided according to their corresponding peaks (1,2,3)

Notes: The first time the column was run, a 10 mL protein sample was used, the second time 40 ml of the protein was used




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