Daniel:Protocols/AmiconUltra: Difference between revisions
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{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | ||
|- style="background-color:#FFCC66;font-size:12pt;font-weight:bold" align="center" | |- style="background-color:#FFCC66;font-size:12pt;font-weight:bold" align="center" | ||
| width=" | | width="120" height="30" | Starting Sample Volume | ||
| width=" | | width="110" | 500 uL dilution | ||
| width=" | | width="110" | Original Buffer Concentration | ||
| width=" | | width="110" | Centrifuge Time | ||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
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| align="center" | 25 | | align="center" | 25 | ||
| align="center" | 0.016% | | align="center" | 0.016% | ||
| align="center" | 15 | | align="center" | 15 min | ||
|} | |} | ||
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==Spin Times and Concentration== | ==Spin Times and Concentration== | ||
[[Image:AmiconSpinTimeTable.png| | [[Image:AmiconSpinTimeTable.png|800px]] |
Latest revision as of 21:52, 13 May 2017
Amicon Ultra Centrifugal Filters[edit]
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[edit]
- 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[edit]
- 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
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 |