User:Tamra L. Fisher/Notebook/Experimental Biological Chem/2012/04/16

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Objective

Remove the His-tag and enterokinase-tag from GFP.

Description

Dr. Hartings very kindly purified the GFP (D1C) that was expressed over the weekend using FPLC and a nickel column. This will allow us to proceed with our project.

GFP Digestion of Tags

  1. Mix 0.8mL of the dialyzed protein with 0.1mL of EKMax™ Buffer, 4μL of EKMax™, and .099mL H2O.
    • See [[1]] for more info on this enzyme.
    • Make four of these reaction mixtures.
  2. Place these mixtures on a heat block at 37°C overnight.



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