Amicon Ultra-0.5 mL Centrifugal Filters for DNA and Protein Purification and Concentration
With their vertical membrane design, Amicon Ultra 0.5 mL filters deliver great performance and lower spin times for protein purification.
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With their vertical membrane design, Amicon Ultra 0.5 mL filters deliver great performance and lower spin times for protein purification. 간단히
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개요
사양
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Documentation
FAQ
| Question | Answer |
|---|---|
| What is the recommended rotor for Amicon Ultra units? | Any rotor capable of accommodating 22.7 cm in diameter and 124 mm length centrifuge tubes for the Amicon Ultra-4 and 35 mm in diameter and 121 mm in length for the Amicon Ultra-15 can be used for Amicon Ultra units. |
| How should I select the appropriate Molecular Weight Cutoff (MWCO) for Amicon Ultra? | Select a Molecular Weight Cutoff that is 2x smaller that the Molecular Weight (MW) of the solute of interest. |
| Which membranes are available in Amicon Ultra units? | Amicon Ultra units are only available with Ultracel regenerated cellulose material. This high recovery/low binding membrane is offered in 3,000, 10,000, 30,000, 50,000 & 100,000 MWCO. |
| Can the Amicon Ultra be spun to dryness? | No, the Amicon Ultra has a built in deadstop. |
| What is the chemical compatibility of Amicon Ultra? | Amicon Ultra units are intended for use with biological fluids and aqueous solutions. Please refer to the user guide, catalogue, or contact Technical Support. |
| Do you have any centrifugal filter units that are pyrogen free? | We do not have any centrifugals devices that we claim are pyrogen free. Even the sterile devices could contain pyrogens. |
관련 제품 및 어플리케이션
관련 제품: Application Facete
| Protein Purification |
| Dialysis and Filtration |
관련 제품: Brand Facete
| Ultracel® |
| Amicon® |
카테고리
| Life Science Research > Protein Sample Preparation > Protein Concentration & Dialysis > Sample Prep Centrifugal Filter Units |
- Concentrates 500 µL down to 15 µL
- Concentration factor 25 x to 30 x
- High sample recovery (greater than 90% of dilute starting solution)
- Fast processing time - 10 to 30 minutes
- Consistent recoveries thanks to the reverse spin capability
- 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-0.5 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 – DNA Concentration
The Amicon Ultra-0.5 device provides the best balance between recovery and spin time for double-stranded DNA for base pairs ranging from 137 to 1159. To achieve maximum PCR product recovery and primer removal with primers greater than 20 bases, one or two additional spins with Tris-EDTA (TE) buffer are recommended.
| Typical Recovery of Nucleotides from Amicon Ultra-0.5 30K Devices | ||||
| PCR Product (base pairs) | PCR Primer (bases) | PCR Recovery (%) | PCR Primer Removal (%) | TE Washes (number) |
| 137 | 10 | ≥95 | ≥90 | 0 |
| 20 | ≥90 | ≥85 | 1 | |
| 48 | ≥90 | ≥75 | 2 | |
| 301 | 10 | ≥90 | ≥90 | 0 |
| 20 | ≥85 | ≥90 | 1 | |
| 48 | ≥90 | ≥80 | 2 | |
| 648 | 10 | ≥95 | ≥90 | 0 |
| 20 | ≥90 | ≥90 | 1 | |
| 48 | ≥95 | ≥90 | 2 | |
| 1159 | 10 | ≥90 | ≥90 | 0 |
| 20 | ≥90 | ≥95 | 1 | |
| 48 | ≥95 | ≥95 | 2 | |
| Spin conditions: 40º fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume, 20-30 µL final volume, 10 minute spin, n=12. | ||||
Performance – Protein Concentration
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Spin Conditions: 40º fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume. Protein markers used: Cytochrome c for 3K and 10K, BSA for 30K and 50K, and IgG for 100K, n=8.
Typical Concentrate Volume/Concentration Factor vs. Spin Time
| Concentrate Volume / Concentration Factor | ||||||||||
| 3K device | 10K device | 30K device | 50K device | 100K device | ||||||
| Spin time (min) | Conc. Volume (µL) | Conc. Factor (x) | Conc. Volume (µL) | Conc. Factor (x) | Conc. Volume (µL) | Conc. Factor (x) | Conc. Volume (µL) | Conc. Factor (x) | Conc. Volume (µL) | Conc. Factor (x) |
| 5 | 215 | 2 | 66 | 7 | 42 | 12 | 28 | 18 | 58 | 9 |
| 10 | 114 | 4 | 35 | 14 | 23 | 22 | 20 | 25 | 19 | 26 |
| 15 | 80 | 6 | 22 | 22 | 19 | 27 | 17 | 30 | 15 | 33 |
| 20 | 62 | 8 | 20 | 24 | 17 | 30 | 15 | 33 | 13 | 36 |
| 30 | 48 | 10 | 15 | 31 | 15 | 32 | 15 | 36 | 11 | 41 |
| Spin Conditions: 40º fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume. Protein markers used: Cytochrome c for 3K and 10K, BSA for 30K and 50K, and IgG for 100K, n=12. Shaded volumes were used for the calculation of protein recovery in the Typical Concentrate Recovery table. | ||||||||||
Typical Retention of Protein Markers
| Marker/Concentration | Molecular Weight | Device NMWL | % Retention | Spin Time (min) |
| α-Chymotrypsinogen (1 mg/mL) | 25,000 | 3K | > 95 | 30 |
| Cytochrome c (0.25 mg/mL) | 12,400 | > 95 | 30 | |
| Vitamin B-12 (0.2 mg/mL) | 1,350 | < 42 | 30 | |
| α-Chymotrypsinogen (1 mg/mL) | 25,000 | 10K | > 95 | 15 |
| Cytochrome c (0.25 mg/mL) | 12,400 | > 95 | 15 | |
| Vitamin B-12 (0.2 mg/mL) | 1,350 | < 42 | 15 | |
| BSA (1 mg/mL) | 67,000 | 30K | > 95 | 10 |
| Ovalbumin (1 mg/mL) | 45,000 | > 95 | 10 | |
| Cytrochorme c (0.25 mg/mL) | 12,400 | < 35 | 10 | |
| BSA (1 mg/mL) | 67,000 | 50K | > 95 | 10 |
| Ovalbumin (1 mg/mL) | 45,000 | ~ 40 | 10 | |
| Cytrochorme c (0.25 mg/mL) | 12,400 | < 20 | 10 | |
| Thyroglobulin (0.5 mg/mL) | 677,000 | 100K | > 95 | 10 |
| IgG (1 mg/mL) | 156,000 | > 95 | 10 | |
| Ovalbumin (1 mg/mL) | 45,000 | < 30 | 10 | |
| Spin Conditions: 40º fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume, n=12. | ||||
Typical Concentrate Recovery
| Marker/Concentration | Molecular Weight | Device NMWL | Spin Time (min) | Concentrate Volume (µL) | Concentration Factor (x) | Concentrate Recovery (%) |
| Cytochrome c (0.25 mg/mL) | 12,400 | 3K | 30 | 48 | 10 | 98 |
| Cytochrome c (0.25 mg/mL) | 12,400 | 10K | 15 | 22 | 22 | 93 |
| BSA (1 mg/mL) | 67,000 | 30K | 10 | 23 | 22 | 97 |
| BSA (1 mg/mL) | 67,000 | 50K | 10 | 20 | 25 | 92 |
| IgG (1 mg/mL) | 156,000 | 100K | 10 | 19 | 26 | 92 |
| Spin Conditions: 40º fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume, n=12. The shaded volumes were taken from the Typical Concentrate Volume/Concentration Factor vs. Spin Time table. | ||||||
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-0.5 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 | 500 |
| Typical final concentration volume, µL | 15 – 20 |
| Recommended relative centrifugal force | 14,000 x g for concentration spin 1,000 x g for reverse spin |
| Maximum relative centrifugal force | 15,000 x g |
| Active membrane area, cm2 | 1 cm² |
| Hold up volume, µL | < 1 |
| Materials | |
| Filter device | Copolymer styrene/butadiene |
| Membrane | Ultracel® low-binding regenerated cellulose |
| Collection tube | Polypropylene |
| The ultrafiltration membranes in Amicon Ultra-0.5 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: 9.4 (0.37) |
| Length: 29.6 (1.17) | |
| Tube, mm (in.) | Diameter: 10.8 mm |
| Length: 43.0 (1.69) | |
| Filter device and tube, mm (in.) | Length (concentration mode; device in tube): 4.99 cm (1.96 in.) |
| Length (reverse spin; device upside down in tube): 37.3 (1.47) | |
Chemical Compatibility of Amicon Ultra Filters
| Acids | Concentration | Acids | Concentration |
| Acetic acid | ≤ 50%* | Phosphoric acid | ≤ 30% |
| Formic acid | ≤ 5%* | Sulfamic acid | ≤ 3% |
| Hydrochloric acid | ≤ 1.0 M | Sulfuric acid | ≤ 3% |
| Lactic acid | ≤ 50% | Trichloroacetic acid (TCA) | ≤ 10%* |
| Nitric acid | ≤ 10% | Trifluoroacetic acid (TFA) | ≤ 30%* |
| Alkalis | Concentration | Alkalis | Concentration |
| Ammonium hydroxide | ≤ 10% | Sodium hydroxide | ≤ 0.5 M |
| Alcohols | Concentration | Alcohols | Concentration |
| n-Butanol | ≤ 70% | Isopropanol | ≤ 70% |
| Ethanol | ≤ 70% | Methanol | ≤ 60% |
| Detergents | Concentration | Detergents | Concentration |
| Alconox® detergent | ≤ 1% | Lubrol® PX detergent | ≤ 0.1% |
| CHAPS detergent | ≤ 0.1% | Nonidet™ P-40 surfactant | ≤ 2% |
| Sodium deoxycholate | ≤ 5% | Triton® X-100 surfactant | ≤ 0.1% |
| Sodium dodecyl sulfate (SDS) | ≤ 0.1% | Tween® 20 surfactant | ≤ 0.1% |
| Terg-A-Zyme® detergent | ≤ 1% | ||
| Organic solvents | Concentration | Organic solvents | Concentration |
| Acetone | not recommended | Ethyl acetate | not recommended |
| Acetonitrile | ≤ 20% | Formaldehyde | ≤ 5% |
| Benzene | not recommended | Pyridine | not recommended |
| Carbon tetrachloride | not recommended | Tetrahydrofuran | not recommended |
| Chloroform | not recommended | Toluene | not recommended |
| Dimethyl sulfoxide (DMSO) | ≤ 5%* | ||
| Miscellaneous | Concentration | Miscellaneous | Concentration |
| Ammonium sulfate | Saturated | Phenol | ≤ 1% |
| Diethyl pyrocarbonate | ≤ 0.2% | Phosphate buffer (pH 8.2) | ≤ 1 M |
| Dithiothreitol (DTT) | ≤ 0.1 M | Polyethylene glycol | ≤ 10% |
| Glycerine | ≤ 70% | Sodium carbonate | ≤ 20% |
| Guanidine HCl | ≤ 6 M | Tris buffer (pH 8.2) | ≤ 1 M |
| Imidazole | ≤ 100 mM | Urea | ≤ 8 M |
| Mercaptoethanol | ≤ 0.1 M | ||
| * Contact with this chemical may cause materials to leach out of the component parts. Solvent blanks are recommended to determine whether leachables represent potential assay interferences. | |||