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Objective
FPLC of MBP-intein.
Description
- Inject about 20mL of the dialyzed supernatent onto the anion-exchange column. (except for the first time when only 5mL was injected)
- Run 25mL of 50mM Tris (pH~9) over the column.
- Start fraction collection. Run a gradient of 50mM Tris from 0M NaCl to 0.5M NaCl over 20 minutes. Collect in 5mL fractions.
- Run 25mL of 50mM Tris + 1M NaCl (pH~9.15) over column.
- Run 25mL of 50mM Tris (pH~9) over the column.
- Repeat until all of the supernatent has been injected and eluted off of the column.
Data
This is the output from the HPLC:
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AU Biomaterials Design Lab
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