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Thermo Scientific™ Pierce™ Protein Concentrators PES, 3K or 5K MWCO, 0.5–100 mL
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Description
Easily and quickly concentrate proteins and biological samples with Thermo Scientific Pierce Protein Concentrators PES (polyethersulfone), which are disposable centrifugation devices that enable fast sample recovery and a short processing time. These concentrator columns can be used with standard microcentrifuge equipment, and samples can be collected without the need for invert spinning. Samples can be concentrated with or without desalting. These products have a molecular weight cutoff (MWCO) of 3K or 5K and can be used for processing volumes of 0.5 to 100 mL.
For protein and other biological samples, such as nucleic acids, with a 3K and 5K MWCO, Pierce Protein Concentrators PES offer rapid protein concentration, excellent sample recovery, and instrument compatibility. These centrifugal devices feature a high-flux polyethersulfone (PES) membrane for concentration, desalting, and buffer exchange of biological samples. These concentrators have MWCOs of 3K and 5K and can be used for processing sample volumes of 0.5–100 mL.
Features of 3K and 5K MWCO Protein Concentrators PES include:
• Rapid protein concentration—sample concentration of 10- to 30-fold can be achieved in 15 or fewer minutes
• Excellent protein recovery—typical protein recovery is >90% (when the sample mass is at least two-fold greater than the posted MWCO)
• Instrument-compatibility—a standard microcentrifuge with swinging or fixed-angle (except 88534) rotors can be used; sample can be collected and recovered without invert spinning
• Versatile—easily perform sample concentration, diafiltration, or buffer-exchange of biological samples
For easy identification, the MWCO value is etched on the side of each device. To assist with sample addition and recovery, a clear window with graduations is located on the side of each device, enabling visual determination of the retentate volume. The vertical design of the membrane allows high-flux rates, low non-specific binding, and negligible protein aggregation on the membrane, giving reliable and consistent results for processing sample volumes between 0.5 and 100 mL. Typical protein recovery is 90% when the mass of the proteins in the sample is at least two-fold greater than the MWCO of the device used. For example, when concentrating IgG (MW ∼150 KDa) samples, use the 5K, 10K, 30K, or 50K MWCO concentrators. Final sample concentrations of 10- to 30-fold can be achieved within 15 minutes.
Protein Concentrators PES applications
• Protein concentration of tissue culture media, antiserum, or monoclonal antibody preparations
• Concentration of protein peaks following gel-permeation chromatography
• Removal of unincorporated label following protein modification reactions
• Concentration and desalt/buffer-exchange after elution of protein from ion-exchange, hydrophobic interaction (HIC), metal chelate, or affinity chromatography columns
• Separation of different sized molecules
Pierce Protein Concentrators PES contain a membrane rated for retaining molecules with a molecular weight at least two-fold greater than the MWCO rating of the PES membrane within the device. Reduced recovery may occur when using a concentrator with molecules smaller than the recommended mass. Protein recovery will vary depending on the specific protein in the sample and its starting concentration.
The dead-stop volume for the 0.5 mL, 2–6 mL, 5–20 mL, and 20–100 mL concentrators is approximately 15 μL, 30 μL, 50 μL, and 350 μL, respectively.
To achieve >90% recovery of protein with the 0.5 mL concentrators, the minimum protein sample concentration should be 0.05 mg/mL. For all other concentrators, the minimum protein sample concentration should be 0.1 mg/mL.
Optimal centrifuges and speeds
The 0.5 mL concentrators can be used effectively at a maximum centrifugal force of 15,000 × g using a fixed angle centrifuge only.
For the 2-6 mL concentrators, swinging-bucket or fixed-angle centrifuge models can be used. Swinging bucket centrifuges capable of generating 2,000–4,000 × g and a maximum centrifugal force of 4,000 × g are recommended. With fixed-angle centrifuges, models capable of generating 5,000–10,000 × g and a maximum centrifugal force of 8,000 × g are recommended.
For the 5-20 mL volume concentrators, swinging-bucket or fixed-angle centrifuge models can be used. Swinging-bucket centrifuges capable of generating 2,000–5,000 × g and a maximum centrifugal force of 4,000 × g are recommended. With fixed-angle centrifuges, models capable of generating 5,000–8,000 × g and a maximum centrifugal force of 6,000 × g are recommended.
For the 20-100 mL concentrators, only swinging-bucket centrifuges capable of generating 1,000–2,000 × g are recommended. Use a maximum centrifugal force of 2,000 × g. These concentrators are not recommended for use in fixed-angle rotors.
Centrifugal force, temperature and sample volume, concentration and viscosity will affect filtration rate and require optimization of time for each application.
Specifications
Specifications
| Content And Storage | Store at room temperature. |
| Description | Pierce™ Protein Concentrators PES, 3K MWCO, 0.5 mL |
| Disposable | Single-use |
| Filter Type | Membrane Filter |
| Format | Microcentrifuge, Spin Column |
| Material (Membrane) | PES |
| MWCO | 3 kDa |
| Sample Volume | ≤0.5 mL |
| Product Line | Pierce |
| Purification Target | Protein |
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Frequently Asked Questions (FAQs)
You may remove excess solvent and smaller moieties by centrifugation through a microconcentrator. This will concentrate your protein sample.
(1) Choose microconcentrator tube (available from a variety of commercial suppliers) with a protein cutoff smaller than the molecular weight of the protein in the sample. Check our Pierce Protein Concentrators PES.
(2) Add 1 µL of 20% w/v SDS to a dry microconcentrator tube (if sample does not already contain SDS).
(3) Slowly add sample (a few microliters at a time) to membrane until membrane is completely wet. Centrifuge to near (but not complete) dryness.
(4) If intention is to desalt sample or buffer exchange: Add ~50 µL water to microconcentrator and spin until nearly dry. Repeat buffer exchange. Sample will remain on membrane. Check our Zeba desalting proteins.
(5) Using a new collection tube, invert membrane and spin at low speed (1000 x g) to elute protein from membrane. Add 2X SDS-Sample Buffer containing 10 mM DTT to membrane: vortex, invert and spin. Final volume should be ~20 µL.
We do not recommend autoclaving the PES protein concentrators. High temperatures will significantly increase the membrane molecular weight cutoff (MWCO). To sterilize, we recommend using a 70% ethanol solution.
Yes, the PES membrane is compatible with desalting and buffer exchange. Protocols for concentrating, desalting, and buffer exchange only are provided in the product manuals.
The protein concentrator columns have an upper compartment that holds the sample, and a lower compartment. The two compartments are separated by a polyethersulfone (PES) semipermeable membrane. Centrifugation is applied to force solvent through the membrane, leaving a more concentrated sample in the upper chamber. The protein concentrator cassettes also contain a PES membrane and a unique channel design that ensures high optimal flux with no optimization effort. Unlike other systems, the fluid is passed parallel to the filter, rather than being pushed through a membrane perpendicularly, which can clog the filter. For maximum protein recovery in both columns and cassettes, samples should have a molecular weight two-fold greater than the molecular weight cutoff (MWCO) of the concentrator membrane.
Pierce polyethersulfone (PES) protein concentrators are offered in six distinct MWCOs of 3K, 5k, 10K, 30K, 50K, and 100K, and the cassettes are offered in five membrane MWCOs of 3K, 10K, 30K, 50K, and 100K. The columns can process four different sample volumes ranges of 100-500 µL, 2-6 mL, 5-20 mL, and 20-100 mL, while the cassettes can process 0.5-6 L. Please see our selection chart and video found here (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-purification-isolation/protein-dialysis-desalting-concentration/protein-concentrators.html).
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