Ultracentrifugation devices for concentration of small-volume protein samples.
The Thermo Scientific Pierce Protein Concentrators PES are disposable ultrafiltration centrifugal devices with a polyethersulfone (PES) membrane for concentration, desalting and buffer exchange of biological samples.
Pierce Protein Concentrators PES can be used for processing sample volumes up to 0.5mL and are available in four distinct molecular-weight cutoffs (MWCOs) of 3K, 10K, 30K and 100K. 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 and provides reliable and consistent results for processing sample volumes between 100 and 500µL. 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. Sample concentration of 10- to 30-fold can be achieved within 10 minutes. The 0.5mL Pierce Concentrators, PES are compatible with most bench-top microcentrifuges with fixed-angle rotors that accommodate 2.2mL tubes.
Rapid protein concentration – sample concentration of 10- to 30-fold can be achieved in 10 minutes or less
Excellent protein recovery – unique design minimizes membrane fouling and provides high speed concentrations, even with particle laden solutions
Instrument-compatible – use standard microcentrifuge with fixed-angle rotors; collect and recover sample without invert spinning
Versatile – easy way to perform sample concentration, diafiltration or buffer-exchange of biological samples
Protein concentration with 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 eluting protein from ion-exchange, hydrophobic interaction (HIC), metal chelate or affinity chromatography columns
Separating large molecules from small molecules
The 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. To achieve >90% recovery of protein, the minimum protein sample concentration should be 0.05mg/mL.
The Pierce Protein Concentrators PES allow accurate control of the dead-stop volume (15µL) and final concentration factor, providing reliable and consistent sample processing. They can be used effectively at a maximum centrifugal force of 15,000 x g at room temperature. Centrifugal force, temperature and sample volume, concentration and viscosity affect filtration rate. Optimize centrifugal time for each application.
Thermo Scientific Pierce Protein Concentrators PES, 10K MWCO provide exceptional recovery of low-concentration samples. Samples of Protein A (~46 kDa) at approximately 0.05mg/mL starting concentrations were centrifuged in Pierce Protein Concentrators PES, 10K MWCO and other suppliers' concentrators at 15,000 x g until a15-fold decrease in sample volume was achieved. Protein recovery was determined by the Thermo Scientific Pierce Micro BCA Protein Assay.
Thermo Scientific Pierce Protein Concentrators PES, 30K MWCO provide exceptional recovery of low-concentration samples. Samples of BSA (~66 kDa) were centrifuged in Pierce Protein Concentrators PES, 30K MWCO and other suppliers' concentrators at 15,000 x g until a 15-fold decrease in sample volume was achieved. Protein recovery was determined by the Thermo Scientific Pierce BCA Protein Assay.
Thermo Scientific Pierce Protein Concentrators PES, 3K MWCO provide exceptional recovery of low-concentration samples. Samples of Aprotinin (~6.5 kDa) were centrifuged in Pierce Protein Concentrators PES, 3K MWCO and other suppliers' concentrators at 15,000 x g until a15-fold decrease in sample volume was achieved. Protein recovery was determined by the Thermo Scientific Pierce BCA Protein Assay.
Separation of polymers from small molecules using Thermo Scientific Pierce Protein Concentrators PES, 100K MWCO. The excipient, sodium starch glycolate (MW > 500,000 to 11,000,000), was separated from a mixture of protease inhibitors leupeptin (MW 475.6), AEBSF (MW 239.69), and bestatin (MW 345) using the Pierce Concentrator PES, 100K MWCO, 0.5mL. The excipient and the inhibitors with exicpient were dissolved in 0.15M NaCl. Half of the volume was loaded into the concentrator after pre-wetting the membrane with 0.15M NaCl. Samples were centrifuged for 10 minutes at 13,000 x g. The flow-through was collected and compared to the non-concentrated controls using reverse-phase HPLC. HPLC conditions: column = Synergi 4u Hydro-RP 80A, Gradient Buffer A = 0.1% TFA in water; Gradient Buffer B = 0.1% TFA in acetonitrile. Panel A. HPLC chromatogram of the excipient and protease inhibitor mixture. Panel B. HPLC chromatogram of the mixture after removal of the excipient using the concentrator. Peaks are identified and labeled.