Desalting Protocol

A follow up on Mimi's post regarding functionalizing your protein, this is the desalting protocol you will use! Note there are two sizes of columns make sure to use 0.5 mL columns!

Desalting Procedure
For maximal protein recovery, add 3 μl of buffer or water above the sample as a stacker. This stacker may be omitted if desired; however, a reduction in protein recovery will result.
A.
Additional Materials Required
•
Variable-speed bench-top microcentrifuge
•
1.5 microcentrifuge collection tubes
B.
Spin Column Preparation
1.
Remove column’s bottom closure and loosen cap (do not remove cap).
2.
Place column in a 1.5-2.0 ml microcentrifuge collection tube and centrifuge at 1,000 × g for 1 minute to remove storage solution.
3.
Blot bottom of column to remove excess liquid.
C.
Sample Loading
1.
Place column in a new collection tube, remove cap and apply 2-12 μl of sample to the top of the compact resin bed.
2.
Apply a 3 μl stacker of ultrapure water or buffer to the top of the resin bed after the sample has absorbed.
3.
Centrifuge at 1,000 × g for 2 minutes to collect the desalted sample. Discard desalting column after use.