Amicon® Ultra-2 mL Centrifugal Filters for DNA and Protein Purification and Concentration
With their vertical membrane design, Amicon® Ultra 2 mL filters deliver great performance and lower spin times
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With their vertical membrane design, Amicon® Ultra 2 mL filters deliver great performance and lower spin times Less
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Overview
Specifications
Ordering Information
Documentation
Related Products & Applications
Related Products By: Application Facete
| Protein Purification |
| Dialysis and Filtration |
Related Products By: Brand Facete
| Ultracel® |
| Amicon® |
Categories
| Life Science Research > Protein Sample Preparation > Protein Concentration & Dialysis > Sample Prep Centrifugal Filter Units |
- Concentrates 2 mL down to 15-70 µl
- 98% sample recovery
- Fast processing time - 10 to 30 minute spin times
- Engineered dead stop
- Reverse spin capability provides consistent recoveries
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Applications
- Concentration of biological samples containing antigens, antibodies, enzymes, nucleic acids, or microorganisms
- Purification of macromolecular components found in tissue culture extracts or cell lysates
- Primer removal, PCR cleanup, removal of linkers or macromolecular labels from a reaction mix, and protein removal prior to HPLC
- Desalting, buffer exchange and protein dialysis
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Material Supplied
The Amicon® Ultra-2 device is supplied with two microcentrifuge tubes. During operation, one tube is used to collect filtrate; the other tube is to recover the concentrated sample.
Performance – Protein Concentration
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Spin conditions: Swinging bucket rotor, 4,000 × g, room temperature, 2 mL starting volume.
Protein markers used: Cytochrome c for 3K and 10K, BSA for 30K and 50K, and IgG for 100K, n=4.
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Spin conditions: 35° fixed angle rotor, 7,500 × g, room temperature, 2 mL starting volume.
Protein markers used: Cytochrome c for 3K and 10K, BSA for 30K and 50K, and IgG for 100K, n=4.
Performance – DNA Concentration
The Amicon® Ultra-2 30K device provides the best balance between PCR recovery and PCR primer removal for double-stranded DNA for base pairs ranging from 137 to 1159.
| Swinging Bucket Rotor | 35° Fixed Angle Rotor | |||||||
| Amicon Ultra Device |
PCR Product (base pairs) |
PCR Primer (bases) | PCR Recovery (%) |
PCR Primer Removal (%) |
Final Volume (μL) |
PCR Recovery (%) | PCR Primer Removal (%) |
Final Volume (μL) |
| 30K | 137 | 10 | 92 | 100 | 37 | 93 | 100 | 27 |
| 20 | 94 | 93 | 39 | 94 | 95 | 31 | ||
| 48 | 95 | 71 | 36 | 93 | 78 | 31 | ||
| 100K | 137 | 10 | 60 | 100 | 33 | 56 | 100 | 16 |
| 20 | 50 | 100 | 33 | 41 | 100 | 17 | ||
| 48 | 50 | 92 | 35 | 39 | 91 | 16 | ||
| Spin conditions: Swinging bucket rotor, 4,000 x g for 40 min; fixed angle rotor, 7,500 x g for 15 min; room temperature, 100 μL PCR diluted to 2,000 μL starting volume, n=3. | ||||||||
Protein Retention and Concentrate Recovery
| Typical Retention of Protein Markers | ||||||
| Marker (Concentration) |
Molecular Weight |
Device NMWL |
% Retention Swinging Bucket |
Spin Time (min) |
% Retention Fixed Angle |
Spin Time (min) |
| Cytochrome c (0.25 mg/mL) | 12,400 | 3K | > 99 | 60 | > 99 | 60 |
| Cytochrome c (0.25 mg/mL) | 12,400 | 10K | > 98 | 40 | > 99 | 20 |
| BSA (1 mg/mL) | 67,000 | 30K | > 99 | 20 | > 99 | 15 |
| BSA (1 mg/mL) | 67,000 | 50K | > 96 | 15 | > 97 | 10 |
| IgG (1 mg/mL) | 156,000 | 100K | > 98 | 30 | > 98 | 20 |
| Spin Conditions: Swinging bucket rotor, 4,000 x g, or 35º fixed angle rotor, 7,500 x g, 2 mL starting volume, room temperature, n=4. | ||||||
Desalting or Diafiltration
Desalting, buffer exchange or diafiltration are important methods for removing salts or solvents in solutions containing biomolecules. The removal of salts or the exchange of buffers can be accomplished in the Amicon® Ultra-2 device by concentrating the sample, then reconstituting the concentrate to the original sample volume with any desired solvent. The process of "washing out" can be repeated until the concentration of the contaminating microsolute has been sufficiently reduced. See example below.
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| Volumes | |
| Maximum initial sample volume, µL | 2.0 |
| Typical final concentration volume, µL | 15 – 70 |
| OPTIMAL and maximum relative centrifugal force – Swinging bucket rotor | 4,000 x g for concentration spin 1,000 x g for recovery spin |
| OPTIMAL and maximum relative centrifugal force – Fixed angle rotor | 7,500 x g for concentration spin 1,000 x g for recovery spin |
| Active membrane area | 1 cm² |
| Hold up volume, µL | < 5 |
| Materials | |
| Filter device | Copolymer styrene/butadiene |
| Membrane | Millipore Ultracel®l ow-binding regenerated cellulose |
| Collection tube | Polypropylene |
| The ultrafiltration membranes in Amicon® Ultra-2 mL devices contain trace amounts of the following: | Amicon® Ultra 3K, 10K, 30K, 50K, 100K devices: glycerine |
| If these materials interfere with analysis, pre-rinse the device with buffer or Milli-Q® water. If interference continues, rinse with 0.1 N NaOH followed by a second spin of buffer or Milli-Q® water. | |
| Dimensions | |
| Filter device, mm (in.) | Diameter: 15.9 (0.63) |
| Length: 70.7 (2.78) | |
| Tube, mm (in.) | Diameter: 13.8 (0.54) |
| Length: 52.9 (2.08) | |
| Filter device and tube, mm (in.) | Length (concentration mode; device in tube): 13.7 (0.54) |
| Length (reverse spin; device upside down in tube): 34.5 (1.36) | |