Daniel:Protocols/AmiconUltra: Difference between revisions
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<li>Add X uL sample and new buffer to 500 uL total volume</li> | |||
<li>Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)</li> | <li>Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)</li> | ||
<li>Discard flow through and add 500 uL new buffer</li> | <li>Discard flow through and add 500 uL new buffer</li> | ||
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===Dialysis Table=== | ===Dialysis Table=== | ||
==Spin Times and Concentration== | ==Spin Times and Concentration== | ||
[[Image:AmiconSpinTimeTable.png|600px]] | [[Image:AmiconSpinTimeTable.png|600px]] |
Revision as of 21:51, 13 May 2017
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:
- CentrifugalFilters-Purification.png
Purification/Concentration
- CentrifugalFilters.png
Buffer Exchange
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