Daniel:Protocols/AmiconUltra

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Amicon Ultra Centrifugal Filters[edit]

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Amicon Order Page/Information

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[edit]

  1. Add X uL sample and buffer to 500 uL total volume
  2. Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
  3. Discard flow through tube and get a new one
  4. Invert filter inside flow through tube and spin down at 14000xg for 1 minute
  5. Alternately, extract sample using pipette

Protocol-Dialysis[edit]

  1. Add X uL sample and new buffer to 500 uL total volume
  2. Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
  3. Discard flow through and add 500 uL new buffer
  4. Repeat steps 2 and 3 as many times as desired

Dialysis Table[edit]

Starting Sample Volume 500 uL dilution Original Buffer Concentration Centrifuge Time
50 10 10% 15 min
20 25 0.4% 15 min
20 25 0.016% 15 min

Spin Times and Concentration[edit]

File:AmiconSpinTimeTable.png