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Amicon^® Ultra Centrifugal Filter Ultrafiltration Guide

Use these four tips to maximize your sample recovery when performing ultrafiltration with Amicon^® Ultra centrifugal filter devices.

Choose the right membrane

A close-up view of a laboratory tube is shown, highlighting the liquid inside. An enlarged section on the right illustrates the separation of particles within the liquid, with arrows indicating movement or flow. The tube has a pink cap at the top.

For regenerated cellulose membrane, choose MWCO that is half the size of your target molecule.

Place device in correct position

Two laboratory tubes are depicted side by side. The left tube has a red "X" mark, indicating an incorrect position, while the right tube has a green checkmark, signifying the correct orientation. A circular component with multiple holes is shown in the background.

Place the device into the centrifuge with one membrane panel parallel with the top of the centrifuge, and the other membrane towards the center of the centrifuge. The panels should not be perpendicular to the outer edge of the centrifuge.

Consider sample sensitivity

A laboratory centrifuge is illustrated, with its lid open, revealing a circular platform with multiple holes for test tubes. Two blue snowflakes are depicted on either side, suggesting a cold environment or refrigeration.

Use refrigerated centrifuge for temperature sensitive samples.

Prevent build-up of protein

An illustration of a laboratory tube with a pink cap is shown. The tube features a blue mechanism that moves up and down, indicating the action of drawing in or dispensing liquid. A pink rectangular component is positioned next to the tube.

Pipette the sample up and down to prevent protein polarization, which can lead to blocked concentration.

HOW TO SELECT AN AMICON^® ULTRA FILTER

Determine the starting volume of your sample
Determine the molecular weight (MW) of your macromolecule of interest, then choose an ultrafiltration membrane with a molecular weight cut-off (MWCO) that is 2-3 times smaller than the MW of your macromolecule.
Select pack size

AMICON^® ULTRA CENTRIFUGAL FILTER DEVICES

Amicon^® Ultra centrifugal filter devices provide fast ultrafiltration with high concentration factors from dilute and complex sample matrices. The vertical design and membrane surface area provide fast sample processing, high sample recovery, and the capability for up to 80-fold concentration factors.

Features

High (90-98%) recovery: Ultracel^® regenerated cellulose membrane in a range of molecular weight cut-offs
Quick processing times: Vertical membrane reduces concentration polarization and membrane fouling for ultra-fast spin times in as little as 10–15 minutes
Low non-specific binding: Heat-sealed membrane minimizes downstream extractables
Convenient sample monitoring: translucent housing and volume gradations
Direct pipetter sample access eliminates processing step to recover concentrate
Reverse spin capability provides consistent and maximum protein recoveries, even with small, dilute samples
Low sample loss: physical deadstop in the filter device prevents spinning to dryness and potential sample loss
Broad chemical compatibility: compatible with a variety of aqueous solutions, pH 1 to 9
Consistent and reliable sample prep: predictable concentration factors, with no drying out of sample
Rapid protein concentration: 25- to 80-fold concentration in a single step

Amicon^®Ultra centrifugal devices are offered with nominal molecular weight limits (NMWLs) of 3 kDa, 10 kDa, 30 kDa, 50 kDa, and 100 kDa in four different volumes:

Amicon^® Ultra 0.5 mL Centrifugal Filters
Amicon^® Ultra 2 mL Centrifugal Filters
Amicon^® Ultra 4 mL Centrifugal Filters
Amicon^®Ultra 15 mL Centrifugal Filters

Molecular Weight (MW)
	Amicon^® Ultra-0.5 Filter	Amicon^® Ultra-2 Filter	Amicon^® Ultra-4 Filter	Amicon^® Ultra-15 Filter
Starting Volume	< 0.5 mL	< 2 mL	<4 mL	< 15 mL
Protein Target Size	MWCO (Da)
6kDa<MW<20kDa	3,000	3,000	3,000	3,000
20kDa<MW<60kDa	10,000	10,000	10,000	10,000
60kDa<MW<100kDa	30,000	30,000	30,000	30,000
100kDa<MW<200kDa	50,000	50,000	50,000	50,000
200kDa<MW	100,000	100,000	100,000	100,000

Choose a Rotor and G-force
	Amicon^® Ultra-0.5 Filter	Amicon^® Ultra-2 Filter	Amicon^® Ultra-4 Filter	Amicon^® Ultra-15 Filter
Starting Volume	< 0.5 mL	< 2 mL	< 4 mL	< 15 mL
Final Volume	15-20 µL	15-70 µL	50 µL	200 µL
Design of Device	Standard 1.5 mL	Standard 15 mL	Standard 15 mL	Standard 50 mL
Fixed Angle (35°) Rotor	14,000 g 1,000 g reverse spin	7,500 g 1,000 g reverse spin	5,000 g for 100k 7,500 g for all other MW	5,000 g
Swinging Bucket Rotor	N/A	4,000 g 1,000 g reverse spin	4,000 g	4,000 g

Concentration Factor
	Amicon^® Ultra-0.5 Filter	Amicon^® Ultra-2 Filter	Amicon^® Ultra-4 Filter	Amicon^® Ultra-15 Filter
Starting Volume	< 0.5 mL	< 2 mL	< 4 mL	< 15 mL
Final Volume	15-20 µL with reverse spin	15-70 µL with reverse spin	50 µL	200 µL
Concentration Factor	25X-30X	14X-67X	80X	75X

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 and protein removal prior to HPLC
Concentration of dilute or pre-purified proteins from column eluents
Desalting, buffer exchange and protein dialysis
Size exclusion and protein fractionation

AMICON^® PRO ADAPTERS

Amicon^® Pro Adapters are designed for use with Amicon^® Ultra centrifugal filtration devices to enable multiple simultaneous protein workflow processes including, but not limited to, dialysis, diafiltration, affinity purification, buffer exchange, filter-aided sample preparation (FASP), and concentration. The Amicon^® Pro Adapter is the only product on the market that offers the flexibility to perform any or all protein workflow manipulations in a singledevice for maximum recovery without sample transfer.The uniquely designed interface between the exchange tube tip and the Amicon^® Ultra device enables greater than 99% buffer exchange in a single spin.

The Amicon^® Pro Adapter is comprised of a 10 mL chamber for wash and buffer exchange steps, a frit that allows the insertion of affinity resins and simultaneous elution and concentration of proteins and antibodies, and an exchange tip that regulates flow for controlled, continuous diafiltration.

Features

Flexibility: designed for simultaneous antibody and protein labelling, purification, sample concentration, or controlled diafiltration
Improved recovery: eliminates sample transfer steps for decreased protein loss
Time savings: combines multiple protein workflow processes in a single device

Applications

Antibody labeling
Buffer exchange with continuous diafiltration

AMICON^® STIRRED CELLS

Amicon^® Stirred Cells use pressurized ultrafiltration (pUF) at high flow rates to process large sample volumes for rapid concentration, diafiltration, and buffer exchange. Gas pressure is applied directly to the cell for filtration. Compatible membrane filters includeUltracel^® RC (regenerated cellulose) andBiomax^® PES (polyethersulfone) membranes, offered in a broad range of nominal molecular weight limits ranging from 1kDato 500kDa. Solutes above the membrane’s nominal molecular weight cut-off (NMWCO) are retained in cell, while water and solutes below the cut-off pass into the filtrate and out of cell.

Features

Large volume ultrafiltration: offered in three different sizes to handle volumes from 50 mL to 400 mL, with an optional external 800 mL reservoir for expanded capacity
Gentle sample concentration: magnetic stirring minimizes concentration polarization and shear denaturation
Rapid ultrafiltration: high flow rates for solutions up to 10%macrosoluteconcentration
Reusable and autoclavable: Amicon^® Stirred Cells can be autoclaved for sterility
Easy, secure setup: quick connectors to tubing
Integrated safety features: with screw threads and a pressure relief valve, there’s no need for external housing. This means easier assembly and disassembly, and very clear confirmation that the device is properly assembled.

Applications

Concentrate, diafilter, and exchange buffers for macromolecule solutions including proteins, enzymes,antibodiesand viruses

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