Amicon Ultra-4 10K Centrifugal Filter Devices: User Guide
Amicon Ultra-4 10K Centrifugal Filter Devices: User Guide
Amicon Ultra-4 10K Centrifugal Filter Devices: User Guide
English
Introduction
Amicon® Ultra-4 10K 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 80-fold concentration.
Typical processing time is 10 to 20 minutes. 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. The concentrate is collected from the filter device sample reservoir using
a pipettor, while the ultrafiltrate is collected in the provided centrifuge tube. The device can be spun in a
swinging-bucket (for optimal performance) or fixed-angle rotor. Amicon® Ultra-4 10K devices are supplied
non-sterile and are for single use only.
Intended Use
The Amicon® Ultra-4 product line includes 5 different cutoffs (Molecular Weight Cutoff, MWCO),
however, the Amicon® Ultra-4 10K device (10,000 MWCO) is the only device intended for in vitro
diagnostic use. It can be used to concentrate serum, urine, cerebrospinal fluid, and other body fluids
prior to analysis. For information on other Amicon® Ultra-4 cutoffs, go to www.millipore.com and enter
Amicon Ultra-4 in the search box.
Materials Supplied
The Amicon® Ultra-4 10K device is supplied with a cap, a filter device, and a centrifuge tube.
Cap
Filter device
Centrifuge tube
Required Equipment
●● Centrifuge with swinging-bucket (preferred) or fixed-angle rotor with wells/carriers that can
accommodate 17 mm × 124 mm 15 mL conical-bottomed tubes
CAUTION: To avoid damage to the device during centrifugation, check clearance before spinning.
●● Pipettor with 200 microliter (μL) tip for concentrate recovery
Suitability
Preliminary recovery and retention studies are suggested to ensure suitability for intended use. See the
“How to Quantify Recoveries” section.
Shelf Life
Shelf life is 3 years from date of manufacture. Refer to expiration date on package label.
Desalting or Diafiltration
Desalting, buffer exchange, or diafiltration are important methods for removing salts or solvents in solutions contain-
ing biomolecules. The removal of salts or the exchange of buffers can be accomplished in the Amicon® Ultra-4 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.
Add 3.95 mL of
10 mM NaCl
or exchange
buffer
Spin to Spin to
concentrate concentrate
4 mL of 50 µL of 4 mL of 50 µL of
1 mg/mL
80 mg/mL 1 mg/mL
80 mg/mL
protein in protein in protein in protein in
100 mM
11.1 mM
NaCl NaCl
3.5
3.0
Filtrate Volume (mL)
2.5
Swinging-bucket rotor
2.0
Fixed-angle rotor
1.5
1.0
0.5
0.0
0 5 10 15 20
Spin time (min)
Spin conditions: Swinging-bucket rotor, 4,000 × g, or fixed-angle rotor, 7,500 × g,
room temperature, 4 mL starting volume. Protein marker used: Cytochrome c, n=6.
Factors that determine sample recovery include the nature of the protein solute relative to the device
MWCO chosen, starting concentration, and concentration factor. Table 3 provides typical recoveries for
the Amicon® Ultra-4 10K device.
Wc × Cc
% concentrate recovery = 100 ×
Wo × Co
Wf × Cf
% filtrate recovery = 100 ×
Wo × Co
Alkalis
Ammonium hydroxide ≤ 10% Sodium hydroxide ≤ 0.5 M
Alcohols
n-Butanol ≤ 70% Isopropanol ≤ 70%
Ethanol ≤ 70% Methanol ≤ 60%
Detergents
Alconox® detergent ≤ 1% Sodium dodecyl sulfate (SDS) ≤ 0.1%
CHAPS detergent ≤ 0.1% Tergazyme® detergent ≤ 1%
Lubrol® PX detergent ≤ 0.1% Triton® X-100 surfactant ≤ 0.1%
Nonidet™ P-40 surfactant ≤ 2% Tween® 20 surfactant ≤ 0.1%
Sodium deoxycholate ≤ 5%
Organic solvents
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
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.
H Use-by date
C CE conformity marking
g Batch code
Initial Final
volume concentrate Qty /
(mL) volume (µL) Product pk 3K 10K 30K 50K 100K
0.5 15–20 Amicon® 8 UFC500308 UFC501008 UFC503008 UFC505008 UFC510008
Ultra-0.5 24 UFC500324 UFC501024 UFC503024 UFC505024 UFC510024
device
96 UFC500396 UFC501096 UFC503096 UFC505096 UFC510096
500 UFC5003BK UFC5010BK UFC5030BK UFC5050BK UFC5100BK
Amicon® Ultra-0.5 Collection Tubes 96 UFC50VL96
2 30–70 Amicon® 24 UFC200324 UFC201024 UFC203024 UFC205024 UFC210024
Ultra-2
device
4 50–100 Amicon® 8 UFC800308 UFC801008* UFC803008 UFC805008 UFC810008
Ultra-4 24 UFC800324 UFC801024* UFC803024 UFC805024 UFC810024
device
96 UFC800396 UFC801096* UFC803096 UFC805096 UFC810096
15 150–300 Amicon® 8 UFC900308 UFC901008* UFC903008 UFC905008 UFC910008
Ultra-15 24 UFC900324 UFC901024* UFC903024 UFC905024 UFC910024
device
96 UFC900396 UFC901096* UFC903096 UFC905096 UFC910096
Technical Assistance
For more information, contact the office nearest you. Up-to-date world-wide contact information is
available on our web site at www.millipore.com/offices. You can also visit the tech service page on our web
site at www.millipore.com/techservice.
Standard Warranty
The applicable warranty for the products listed in this publication may be found at
www.millipore.com/terms (within the “Terms and Conditions of Sale” applicable to your purchase
transaction).
Made in Ireland
M Merck Millipore Ltd. C
Tullagreen,
Carrigtwohill,
Co. Cork, IRL
The M logo, Millipore, Amicon, Milli-Q, and Ultracel are registered trademarks of Merck KGaA, Darmstadt, Germany.
All trademarks of third parties are the property of their respective owners.
© 2014 EMD Millipore Corporation. Billerica, MA, U.S.A. All rights reserved.
PR04316TR, Rev. B, English, 07/14