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Summary: Benjamin Costantino 7/25/04
E63-1 x HIS Protein Purification
Stock Solutions:
LB
100mM IPTG (2.38g IPTG in 100mL ddH2O; filter sterilize; store -20° C)
Ampicillin or Carbenicilllin (use at 50ug/mL)
Solutions Provided From His-Bind Buffer Kit (Novagen Cat.#69755-1):
· 8x Binding Buffer
· 8x Wash Buffer
· 4x Elute Buffer
· 4x Strip Buffer
· 8x Charge Buffer
Other Novagen Products Used:
· Pre packed His- Bind Column (#70971) stored at 4° C
· Bug Buster (#70584) stored Room Temp.
· Benzonase (#70746) stored at -20° C
Inducing Recombinant Protein Expression and Preparing Cell Lysate
A. Induction
1. Streak Amp-LB plate with freezer stock E63-1xHis BL21 cells and incubate plate
overnight
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