Daniel:Protocols/AmiconUltra
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Amicon Ultra Centrifugal Filters
Amicon Ultra Centrifugal filters are microcentrifuge filters that allow you to purify and buffer exchange various MW cutoff biomacromolecules. They use a MW cutoff filter, say 10 kDa, to allow dialysate through while not passing the biomacromolecules. They can be used for buffer exchange via the following general protocol:
Protocol-Filtration
- Add X uL sample and buffer to 500 uL total volume
- Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
- Discard flow through tube and get a new one
- Invert filter inside flow through tube and spin down at 14000xg for 1 minute
- Alternately, extract sample using pipette
Protocol-Dialysis
- Add X uL sample and new buffer to 500 uL total volume
- Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
- Discard flow through and add 500 uL new buffer
- Repeat steps 2 and 3 as many times as desired