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Amicon Ultra-0.5 mL Centrifugal Filters for Protein Purification and Concentration

With their vertical membrane design, Amicon Ultra 0.5 mL filters deliver great performance and lower spin times.
 

  • 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

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
  • Size exclusion and protein fractionation

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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 – Protein Concentration

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.

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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