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How do I prepare a spin column?
Posted April 2, 2019
I'd like to use a 1 mL spin column, such as a Sephadex® G-25, BioGel® P-6 DG Media or other desalting column to purify 100 µL of a reaction mixture. Can you provide a sample protocol?
Answer
Below is a sample procedure on how to set up and use a 1 mL spin column for purification of 100 µL test conjugate:
- Fill with beads
- Drain the solution, top dry
- Buffer exchange using ~4 mL PBS buffer (pH 7.2 - 7.4)
- Spin column at 2000X for 2 minutes
- Exchange current 15 mL tube for a brand new one
- Load sample mixture directly to the top of the desalting column
- Add 10 µL of PBS buffer (pH 7.2 - 7.4)
- Spin column at 2000X for 5 minutes
- Collect elution
- Measure the concentration using a spectrophotomer
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