Amicon® Ultra 2 mL Centrifugal Filters

 Amicon® Ultra 2mL for Protein Purification and Concentration

Amicon® Ultra-2 mL centrifugal filter devices provide fast ultrafiltration, with the capability for high concentration factors and easy concentrate recovery from dilute and complex sample matrices. The vertical design and available membrane surface area provide fast sample processing, high sample recovery (typically greater than 90% of dilute starting solution), and the capability for 50-fold concentration. Solute polarization and subsequent fouling of the membrane are minimized by the vertical design, and a physical deadstop in the filter device prevents spinning to dryness and potential sample loss. Efficient recovery of the concentrated sample (retained species) is achieved by a convenient reverse spin step after collecting the filtrate.

The device can be spun in a swinging bucket or fixed angle rotor. Amicon® Ultra-2 mL devices are supplied non-sterile and are for single use only. With their vertical membrane design and reverse spin, Amicon® Ultra 2 mL filters deliver the highest recovery with shortest spin times.

 

 Advantages

  • 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

 

 Performance

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.

 

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.

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 35° fixed angle rotor, 7,500 x g, 2 mL starting volume, room temperature, n=4. 

 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

 

 DNA Concentration

The Amicon® Ultra-2 mL 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. 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.

DNA Concentration
 

  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.


 Materials