Amicon® Ultra-15 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 15 to 40 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 or fixed-angle rotor. Amicon® Ultra-15 10K devices are supplied non-sterile and are for single use only.
The Amicon® Ultra-15 product line includes 5 different cutoffs (Molecular Weight Cutoff, MWCO), however, the Amicon® Ultra-15 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-15 cutoffs, go to www.millipore。。ｃｏｍ and enter Amicon Ultra-15in the search box.
●Concentration of biological samples containing antigens, antibodies, enzymes, nucleic acids (DNA/RNA samples, either single- or double-stranded), microorganisms, column eluates, and purified samples
●Purification of macromolecular components found in tissue culture extracts and cell lysates, removal of primer, linkers, or molecular labels from a reaction mix, and protein removal prior to HPLC
●Desalting, buffer exchange, or diafiltration
The Amicon® Ultra-15 10K device is supplied with a cap, a filter device, and a centrifuge tube
●Centrifuge with swinging-bucket or fixed-angle rotor with wells/carriers that can accommodate 50 mL tubes
CAUTION: To avoid damage to the device during centrifugation, check clearance before spinning.
●Pipettor with 200 microliter (μL) tip for concentrate recovery
Preliminary recovery and retention studies are suggested to ensure suitability for intended use. See the “How to Quantify Recoveries” section.
Shelf life is 3 years from date of manufacture. Refer to expiration date on package label.
Rinsing Before Use
The ultrafiltration membranes in Amicon® Ultra-15 10K devices contain trace amounts of glycerine. If this material interferes with analysis, rinse the device with buffer or Milli-Q® water before use. If interference continues, rinse with 0.1 N NaOH followed by a second spin of buffer or Milli-Q® water.
CAUTION: Do not allow the membrane in Amicon® Ultra filter devices to dry out once wet. If you are not using the device immediately after rinsing, leave fluid on the membrane until the device is used.
How to Use Amicon® Ultra-15 Centrifugal Filter Devices
1. Add up to 15 mL of sample (12 mL if using a fixed-angle rotor) to the Amicon® Ultra filter device.
2. Place capped filter device into centrifuge rotor; counterbalance with a similar device.
3. When using a swinging-bucket rotor, spin the device at 4,000 × g maximum for approximately 15–40 minutes.When using a fixed-angle rotor, orient the device with the membrane panel facing up and spin at 5,000 × g maximum for approximately 15–40 minutes.NOTE: Refer to Figure 1 and Table 1 for typical spin times.
4. To recover the concentrated solute, insert a pipettor into the bottom of the filter device and withdraw the sample using a side-to-side sweeping motion to ensure total recovery. The ultrafiltrate can be stored in the centrifuge tube.
NOTE: For optimal recovery, remove concentrated sample immediately after centrifugation.
Desalting or Diafiltration
Desalting, buffer exchange, or diafiltration are important methods for removing salts or solvents in solutions containing biomolecules. The removal of salts or the exchange of buffers can be accomplished in the Amicon® Ultra-15 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