Amicon® Ultra 0.5 mL Centrifugal Filters

 Amicon® Ultra 0.5mL Filters for Protein Purification and Concentration

Amicon® Ultra-0.5 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 30-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.

Amicon® Ultra-0.5 devices are supplied non-sterile and are for single use only. With their vertical membrane design and reverse spin, Amicon® Ultra 0.5 mL filters deliver great performance and lower spin times.

Order Amicon® Ultra 0.5 mL filters 
Centrifuge and Rotor Recommendations

 

 Advantages

  • Concentrates 500 µL down to 15 µL
  • Concentration factor 25x to 30x
  • High sample recovery (greater than 90% of dilute starting solution)
  • Fast processing time: 10 to 30 minutes
  • Reverse spin capability provides consistent recoveries

Typical Retention of Protein Markers
 

Device NMWL Marker/Concentration Molecular Weight % Retention Spin Time (min)
3K α-Chymotrypsinogen (1 mg/mL) 25,000 > 95 30
Cytochrome c (0.25 mg/mL) 12,400 > 95 30
Vitamin B-12 (0.2 mg/mL) 1,350 < 42 30
10K α-Chymotrypsinogen (1 mg/mL) 25,000 > 95 15
Cytochrome c (0.25 mg/mL) 12,400 > 95 15
Vitamin B-12 (0.2 mg/mL) 1,350 < 23 15
30K BSA (1 mg/mL) 67,000 > 95 10
Ovalbumin (1 mg/mL) 45,000 > 95 10
Cytrochorme c (0.25 mg/mL) 12,400 < 35 10
50K BSA (1 mg/mL) 67,000 > 95 10
Ovalbumin (1 mg/mL) 45,000 ~40 10
Cytrochorme c (0.25 mg/mL) 12,400 < 20 10
100K Thyroglobulin (0.5 mg/mL) 677,000 > 95 10
IgG (1 mg/mL) 156,000 > 95 10
Ovalbumin (1 mg/mL) 45,000 < 30 10
Spin Conditions: 40° fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume, n=12.


 Amicon® 0.5mL Filter for DNA Purification and Concentration

Performance – DNA Concentration
The Amicon® Ultra 0.5 device provides the best balance between recovery and spin time for double stranded DNA base pairs form 137 to 1150. 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-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: 40° fixed angle rotor, 14,000 x g, room temperature, 500 µL starting volume, 20-30 µL final volume, 10 minute spin, n=12

 

 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, linkers, or macromelecular labels from a reaction mix, and protein removal prior to HPLC
  • Desalting, buffer exchange, and protein dialysis

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: 40° 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 Concentrate Recovery Spin Conditions
 

Marker/Concentration Molecular Weight Device NMWL Spin Time (min) Conc. Volume (µL) Conc. 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: 40° 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. 


 Materials