Amicon Ultra-15 Centrifugal Filter Devicesfor volumes up to 15 mLUser GuideAmicon Ultra-15 10K device for in vitro diagnostic useAmicon Ultra-15 3K, 30K, 50K, and 100K devices for research use only; not for use in diagnostic procedures PR02842_RevB.indd 2910/30/2009 11:42:29 AMNotice The information in this document is subject to change without notice and should not be construed as a commitment by Millipore Corporation or an affiliated corporation. Neither Millipore Corporation nor any of its affiliated corporations assumes responsibility for any errors that may appear in this document. This manual is believed to be complete and accurate at the time of publication. In no event shall Millipore Corporation be liable for incidental or consequential damages in connection with or arising from the use of this manual. 2009 MILLIPORE CORPORATION. ALL RIGHTS RESERVED.PR02842 Rev. B, 10/09Millipore, Amicon, Centricon, Milli-Q, MultiScreen, and Ultracel are registered trademarks of Millipore Corporation. The M mark is a trademark of Millipore Corporation. Alconox and Terg-A-Zyme are registered trademarks of Alconox, Inc. Lubrol is a registered trademark of Imperial Chemical Industries PLC. Nonidet is a trademark of Royal Dutch/Shell Group. Triton is a registered trademark of Union Carbide Corporation. Tween is a registered trademark of ICI Americas, Inc.PR02842_RevB.indd 3010/30/2009 11:42:29 AMContentsIntroduction 1 Applications .2 Materials Supplied .3 Required Equipment 4 Suitability .4 Device Storage 4 Specifications 5 Chemical Compatibility 7 How to Use Amicon Ultra-15 Centrifugal Filter Devices .9 How to Quantitate Recoveries .10 Performance - Protein Concentration 13 Flow Rate.13 Protein Retention and Concentrate Recovery18 Maximizing Sample Recovery .21 Desalting or Diafiltration .22 Centrifugal Product Ordering Information 23 Technical Assistance .24 In Vitro Diagnostic Product Labeling 25 Standard Warranty .26PR02842_RevB.indd 3110/30/2009 11:42:29 AMIntroductionMillipores Amicon Ultra-15 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 60 minutes depending on Nominal Molecular Weight Limit (NMWL). 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 devices are supplied non-sterile and are for single use only.PR02842_RevB.indd 110/30/2009 11:42:26 AM2www.millipore.comIntroduction, continuedThe Amicon Ultra-15 product line includes 5 different cutoffs (Nominal Molecular Weight Limit, NMWL, or Molecular Weight Cutoff, MWCO): Amicon Ultra 3K device 3,000 NMWL Amicon Ultra 10K device 10,000 NMWL Amicon Ultra 30K device 30,000 NMWL Amicon Ultra 50K device 50,000 NMWL Amicon Ultra 100K device 100,000 NMWL Amicon Ultra-15 10K filter devices are for in vitro diagnostic use and can be used to concentrate serum, urine, cerebrospinal fluid, and other body fluids prior to analysis. Amicon Ultra-15 3K, 30K, 50K, and 100K filter devices are for research use only and not for use in diagnostic procedures. ApplicationsConcentration of biological samples containing antigens, antibodies, enzymes, nucleic acids (DNA/RNA samples, either single- or double-stranded), microorgan- isms, column eluates, and purified samplesPR02842_RevB.indd 210/30/2009 11:42:26 AMAmicon Ultra-15 Centrifugal Filter Devices3Applications, continuedPurification 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 Materials SuppliedFilter deviceCentrifuge tubeCap The Amicon Ultra-15 device is supplied with a cap, a filter device, and a centrifuge tube.PR02842_RevB.indd 310/30/2009 11:42:27 AM4www.millipore.comRequired EquipmentCentrifuge with swinging bucket or fixed angle rotor with wells/carriers that can accommodate 50 mL tubesCAUTION: To avoid damage to the device during centrifugation, check clearance before spinning. Pipettor with 200 microliter (L) tip for concentrate recovery SuitabilityPreliminary recovery and retention studies are suggested to ensure su