Product Data AcroPrep™ Membrane-bottom Filter Plates Multi-well, Membrane-bottom Plates for High Throughput Sample Preparation and Detection Procedures • Broad selection – available in 96- or 384-well formats with a variety of membranes, plate color choices, well volumes, and outlet tip lengths for use in a multitude of sample preparation and detection processes. • Automation friendly – designed in accordance with the ANSI/Society for Biomolecular Screening (SBS) standards. Rigid construction enables the plate to be easily maneuvered by robotic equipment and assures that plates do not flex and jam detectors or other instrumentation. • Low binding, chemically resistant – polypropylene housing assembly is compatible with a broad range of aqueous and organic liquids, and is noted for its low biomolecule binding so that non-specific adsorption to the plate is minimized. • Serialized barcodes – serialized barcode labels allow for the use of automated tracking systems. • No crosstalk – specially engineered fluid deflectors and outlet tips on the bottom of the plate were designed to reduce the potential for crosstalk. Elimination of crosstalk is assured through individually sealing a membrane in each well using proprietary sealing technology. Applications • • • • • • • • • • Sample prep/gross fractionation DNA purification, concentration, and fractionation Receptor:ligand binding assays Prefiltration Clarification of cell lysates and tissue homogenates Preparation of proteinaceous solutions Drug binding studies Low protein or nucleic acid binding procedures Chemiluminescence detection Radioactive (isotopic) detection • • • • • • • • Fluorescence detection Combinatorial chemistry library screening Cleavage assays Bead-based assays Protein purification, concentration, and fractionation Capture from liquid chromatography Antibody purification Contaminant removal such as DNA, RNA, or other charged molecules Plate Selection Low Biomolecule Binding Solvent Resistance Required Hydrophobic Membrane (Solvent Based Solutions) PTFE Membrane Applications • Molecular Synthesis Reactions • Drug Synthesis Reactions • Resin Cleavage Reactions • Combi-chem Library Screening • Sample Prep Involving Solvents 96 Well 350 µL 1 mL 0.2 µm 0.45 µm 0.2 µm 0.45 µm Natural Housing Natural Housing Natural Housing Natural Housing PN 5037 PN 5038 PN 5055 PN 5056 Low Biomolecule Binding Solvent Resistance Not Required Integrated Prefilter Polypropylene Prefilter Applications • Debris Removal • Low Salt DNA Purification • Rapid Plasmid Preparation • Proteinaceous Sample Purification • Tissue Homgenates Glass Fiber Prefilter Applications • Plasmid Preparation • Lysate Clearance • Protein Solutions 96 Well 96 Well 1 mL 350 µL 350 µL Polypropylene/1.2 µm Supor® Membrane 3.0 µm GF/0.2 µm Bio-Inert® Membrane 3.0 µm GF/0.2 µm Bio-Inert Membrane PN 5041 PN 5046 PN 5053 Hydrophilic Membrane (Aqueous or Solvent Based Solutions) GHP Membrane (Hydrophilic Polypropylene) Applications • Cleavage Reactions • In-gel Digestion • Combi-chem Library Screening • Drug Binding Studies • Sample Prep • Bead/Resin Applications 384 Well 96 Well 350 µL 0.2 µm 1 mL 0.2 µm 0.45 µm 0.45 µm Natural Housing Natural Housing Natural Housing PN 5052 PN 5054 0.45 µm Natural Housing PN 5045 Natural Housing White Housing Black Housing PN 5030 PN 5043 PN 5044 100 µL Short Tip Long Tip PN 5071 PN 5070 No Prefilter Polyester Screen Applications • Sieve Filtration • Cytometry Sample Preparation Bio-Inert Membrane Supor® Membrane GHP Membrane (Hydrophilic Nylon) (Modified PES) (Hydrophilic Polypropylene) Applications • Sample Prep • Drug Binding Studies • Purification of Oligos from Polyacrylamide Gels • Bead Assays and Applications • Resin-based Purifications Applications • Sample Prep • Drug Binding Studies • Purification of Oligos from Polyacrylamide Gels • Resin-based Purifications • Lysate Clearance Applications • Cleavage Reactions • In-gel Digestion • Combi-chem Library Screening • Drug Binding Studies • Sample Prep • Bead/Resin Applications 96 Well 96 Well 96 Well 96 Well 384 Well 350 µL 350 µL 350 µL 1 mL 100 µL 30-40 µm 0.2 µm 1.2 µm 1.2 µm PN 7267 PN 5042 0.45 µm 1.2 µm PN 5029 PN 5039, 5039L PN 5065 See Above Short Tip Long Tip PN 5085 PN 5084 www.pall.com/lab Plate Selection High Biomolecule Binding Binding Media Glass Fiber Applications • Receptor Ligand Binding Studies • Cell-based Assays • Sample Prep • Gross Fractionation 96 Well 384 Well 1 mL 350 µL 1.0 µm Natural Housing PN 5031, 5031L 100 µL 1.0 µm 1.0 µm 1.0 µm 1.0 µm White Housing Long Tip Natural Housing White Housing PN 5032 PN 5051 Short Tip Long Tip Short Tip Long Tip PN 5073 PN 5072 PN 5073W PN 5072W Ion Exchange Separations Membrane Chromatography Mustang™ Membrane Applications • Fractionation of Proteins/Peptides • Capture from Liquid Chromatography • Antibody Purification • Contaminant Removal Such as DNA, RNA, or Other Charged Molecules 96 Well 1 mL 350 µL Mustang S (Cation Exchange) Mustang Q (Anion Exchange) Mustang S (Cation Exchange) Mustang Q (Anion Exchange) PN 5048 PN 5047 PN 5063 PN 5062 Molecular Separations Ultrafiltration Polyethersulfone Membrane (OmegaTM Membrane) Applications • Concentrate, Desalt, and Purify DNA, RNA, Oligos, Proteins, and Peptides • Size Exclusion Applications • Clean-up of Labeling Reactions • Determination of Free vs. Bound Drug in Plasma • Purification of Nucleic Acids and Proteins 96 Well 350 µL 1 mL 3K 384 Well 100 µL 3K 10K 30K 100K PN 5033 PN 5034 PN 5035 PN 5036 10K 30K 100K Long Tip Short Tip Long Tip Short Tip Long Tip Short Tip Long Tip PN 5066 PN 5077 PN 5076 PN 5079 PN 5078 PN 5081 PN 5080 Plate Color Selection Guide Detection results can be further optimized by selecting the proper plate color. Use the following recommendations as a guideline for easy selection. Natural (semi-opaque) – fluorescence, TRF White (opaque) – radioactivity, chemiluminescence Black (opaque) – lowest background for fluorescence, TRF www.pall.com/lab Specifications AcroPrep™ 96 Filter Plates, 350 µL Wells Materials of Construction Housing: Polypropylene (natural, white, or black) Lid: Polystyrene Filter Media: 0.2 and 0.45 µm GHP membrane (hydrophilic polypropylene) 0.2 and 0.45 µm PTFE membrane (hydrophobic PTFE on polyester support) 3.0 µm Glass Fiber/0.2 µm Bio-Inert® membrane (modified hydrophilic nylon) 0.2 µm Bio-Inert membrane (modified hydrophilic nylon) 1.0 µm (nominal) Glass Fiber (borosilicate glass without binder over polypropylene support) 3, 10, 30, and 100K Omega™ membrane (modified polyethersulfone) 0.45 and 1.2 µm Supor® membrane (hydrophilic polyethersulfone) 0.8 µm Mustang™ Q and S membranes (hydrophilic polyethersulfone) 30-40 µm polyester screen AcroPrep 96 Well Plates, 1 mL Wells Materials of Construction Housing: Polypropylene (natural) Filter Media: 0.2 and 0.45 µm GHP membrane (hydrophilic polypropylene) 0.2 and 0.45 µm PTFE membrane (hydrophobic PTFE on polyester support) 3.0 µm Glass Fiber/0.2 µm Bio-Inert membrane (modified hydrophilic nylon) 1 µm (nominal) Glass Fiber (borosilicate glass without binder over polypropylene support) 0.8 µm Mustang Q and S membranes (hydrophilic polyethersulfone) 3K Omega membrane (modified polyethersulfone) 1.2 µm Supor membrane (hydrophilic polyethersulfone) Dimensions Length: 12.78 cm (5.03 in.) Width: 8.55 cm (3.37 in.) Height (without lid): 1.44 cm (0.57 in.) Height (with lid): 1.66 cm (0.66 in.) Well-bottom Area: 0.25 cm2 Maximum Well Volume 380 µL Recommended Working Volume 300 µL Recommended Operating Vacuum 25.4 cm Hg (10 in. Hg) or greater Filtration by Centrifugation 500-3,000 x g Typical Sample Hold-up Volume < 15 µL/well Dimensions Length: 12.78 cm (5.03 in.) Width: 8.55 cm (3.37 in.) Height (without lid): 2.87 cm (1.13 in.) Tip Length: 0.53 cm (0.21 in.) Well-bottom area: 0.25 cm2 Maximum Well Volume 1 mL Recommended Working Volume 900 µL Recommended Operating Vacuum 25.4 cm Hg (10 in. Hg) or greater Filtration by Centrifugation 1,000-3,000 x g Typical Sample Hold-up Volume < 27 µL/well AcroPrep 384 Well Plates, 100 µL Wells Materials of Construction Housing: Polypropylene (natural) Lid: Polystyrene Media: 0.45 µm GHP membrane (hydrophilic polypropylene) 1 µm (nominal) Glass Fiber (borosilicate glass without binder over polypropylene support) 1.2 µm Supor (hydrophilic polyethersulfone) 10, 30, and 100K Omega ultrafiltration membrane (modified polyethersulfone) Dimensions Length: 12.78 cm (5.03 in.) Width: 8.55 cm (3.37 in.) Height (with lid): 1.66 cm (0.66 in.) Height (without lid): 1.44 cm (0.57 in.) Long Tip Length: 0.41 cm (0.16 in.) Total Plate Height Including Long Tips: 1.85 cm (0.73 in.) Well-bottom Area: 0.05 cm2 Maximum Well Volume 100 µL Recommended Working Volume 80 µL Recommended Operating Vacuum 50.8 cm Hg (20 in. Hg) or greater Filtration by Centrifugation 500-3,000 x g Typical Sample Hold-up Volume < 4 µL per well Methodology Filtration vs. Retention 1. Place filter plate on vacuum manifold or hold the plate so that the outlet tips are not touched. 3a. Release vacuum from the manifold. Remove filter plate and retained sample for further processing. 2. Add sample and incubate. Apply vacuum. OR 3b. Release vacuum from the manifold. Remove filter plate. Remove collection (receiver) plate and utilize collected filtrate in downstream applications. AcroPrep™ Filter Plate Operating Instructions AcroPrep filter plates are ideally suited for both manual use and high-volume robotics applications. Manual Vacuum Filtration Procedures 1. During use, hold the plate so that the outlets on the bottom of the plate are not touched. This prevents liquid flow due to wicking. 2. Add the solution to be filtered and incubate. Place the plate on the vacuum manifold and apply vacuum. (See product specifications for recommended vacuum.) 3. Release vacuum from the manifold. DO NOT RELEASE VACUUM BY PULLING THE CORNER OF THE PLATE AS IT WILL DEGRADE THE MANIFOLD GASKET. Robotic Vacuum Filtration Procedure 1. AcroPrep filter plates can be stacked with or without lids in place. 2. Process the plates in the robotic equipment in the same way as a standard solid-bottom plate. 3. Transfer the AcroPrep filter plates to a vacuum manifold station and apply the vacuum. (See product specifications for recommended vacuum.) 4. Gently tap the plate to remove any hanging droplets. 5. Add wash solution(s) and filter as needed. www.pall.com/lab Methodology (continued) Compatible Robotic Systems AcroPrep™ filter plates offer a robust platform for sample preparation and detection applications and are compatible with a variety of currently available robotic systems. Following is a short list of just some of the compatible equipment. Liquid Handlers • Adept Technology, Inc. • Beckman Technologies • Cyber Lab • Hamilton Company • Matrix Technologies Corp. • Oyster Bay Pump Works, Inc. • PerkinElmer Inc. • Porvair • Qiagen, Inc • Robbins Scientific Corp. • Robocon • Tecan U.S. • Titantele • Tomtec • Velocity 11 • Waters Vacuum Manifolds • Pall Life Sciences • 3M • Innovative Microplates • Millipore • Porvair • Qiagen, Inc. • Whatman Note: Polypropylene is an inert material with low biomolecule binding. This is important to prevent nonspecific binding. If a small amount of binding still occurs, classical passivation procedures can be used (see Nanosep® Application Guide – PN 32989). Performance Figure 1 Lower Level of False Positives with AcroPrep 96 Filter Plates Than Competitive Plates Pall AcroPrep 96 Competitor M 3 µg/mL PE-Ab 47 24 32 31 55 30 29 42 10 14 34 42 52 29 45 14 101 1313 678 1425 826 752 1118 1300 1728 692 743 905 889 770 403 895 Avg. SD % CV 33 13 40.97 908 399 43.89 Pall AcroPrep 96 Competitor M Pall AcroPrep 96 Competitor M 4 µg/mL PE-Ab 58 74 65 126 54 175 38 41 77 108 88 215 168 99 231 179 191 2234 972 1467 894 806 1064 1663 1302 1149 1229 1957 1012 2239 1919 1177 5 µg/mL PE-Ab 172 209 86 120 119 52 177 108 1307 1194 937 1170 455 696 478 17 1187 1740 2146 1084 4021 1628 2072 767 3584 2842 319 2657 288 656 1053 869 Avg. SD % CV 112 63 55.98 1330 556 41.84 Avg. SD % CV 456 457 100.18 1682 1129 67.11 The serological immunoassays were performed with two manufacturer’s 96 well filter plates to compare Competitor M filter plates to Pall AcroPrep 96 filter plates with 1.2 µm Supor® membrane. The serological immunoassays were performed using a single serum sample and three concentrations of rPhycoerythrin conjugated antibody. The results from these filter plates were read with one Luminex LX100 Instrument. The results presented are for the sixteen individual responses, the average, the standard deviation, and the percent coefficient of variation for the single serum sample. The Pall Life Sciences AcroPrep 96 filter plates exhibit a lower level of false positive results than Competitor M filter plates. Performance (continued) Figure 2 AcroPrep™ 96 Filter Plates with Prefilter Speeds Sample Processing Times 12 5042 5046 Flow Rate (mL/sec) 10 COMP 8 6 4 2 0 100 µL 200 µL Volume Filtered The use of a prefilter shortens filtration times and allows greater flow rates to be maintained at higher loads. Average flow rates for 100 and 200 µL lysate samples were calculated for the AcroPrep 96 filter plate with Bio-Inert® membrane (PN 5042), AcroPrep 96 filter plate with glass fiber over Bio-Inert membrane (PN 5046), and Competitor hydrophilic PVDF plate (COMP). Error bar indicates standard error (n = 8). See Pall Corp. Technical Report PN 33308 for complete experimental details. Figure 3 AcroPrep 96 UF Filter Plates Remove Salts While Maintaining High Sample Recoveries AcroPrep 96 30K AcroPrep 96 10K 100 99.1 97.3 94 93 100 90 90 80 80 70 70 60 60 50 40 50 40 30 30 20 20 10 10 0 99.2 98.1 94 95 0 % Salt Removed % Protein Recovered % Salt Removed Ovalbumin (45kDa) 0.1 mg/mL 500 mM NaCl BSA (66kDa) 0.1 mg/mL 500 mM NaCl Ovalbumin (45kDa) 1.0 mg/mL 200 mM NaCl BSA (66kDa) 1.0 mg/mL 200 mM NaCl % Protein Recovered In addition to desalting, ultrafiltration is also useful for the concentration of biomolecules. The efficiency of AcroPrep 96 UF filter plates with Omega™ membrane to remove salts and other small molecules is not at the expense of sample recovery. Using the AcroPrep 96 10K filter plate, recovery of ovalbumin proteins were greater than 90% (n = 3) with salt removal efficiencies at greater than 95%. Using BSA and a 30K MWCO membrane, similiar results were observed. A representative experiment is shown. See Pall Corp. Technical Report PN 33309 for complete experimental details. www.pall.com/lab Ordering Information AcroPrep™ 96 Filter Plates, 350 µL well AcroPrep 384 Filter Plates, 100 µL well Part Number Description Pkg Part Number Description Pkg 5033 3K Omega™ membrane, natural 10/pack 5076 10K Omega membrane, long tips, natural 10/pack 5034 10K Omega membrane, natural 10/pack 5077 10K Omega membrane, short tips, natural 10/pack 30K Omega membrane, long tips, natural 10/pack 5035 30K Omega membrane, natural 10/pack 5078 5036 100K Omega membrane, natural 10/pack 5079 30K Omega membrane, short tips, natural 10/pack 5045 0.2 µm GHP membrane, natural 10/pack 5080 100K Omega membrane, long tips, natural 10/pack 5030 0.45 µm GHP membrane, natural 10/pack 5081 100K Omega membrane, short tips, natural 10/pack 0.45 µm GHP membrane, long tips, natural 10/pack 10/pack 5043 0.45 µm GHP membrane, white 10/pack 5070 5044 0.45 µm GHP membrane, black 10/pack 5071 0.45 µm GHP membrane, short tips, natural 5037 0.2 µm PTFE membrane, natural †† 10/pack 5072 1.0 µm glass fiber media, long tips, natural 10/pack 5072W 1.0 µm glass fiber media, long tips, white 10/pack 5073 1.0 µm glass fiber media, short tips, natural 10/pack 5073W 1.0 µm glass fiber media, short tips, white 10/pack 5084 1.2 µm Supor membrane, long tips, natural 10/pack 5085 1.2 µm Supor membrane, short tips, natural 10/pack 5038 0.45 µm PTFE membrane, natural †† 10/pack 5042 0.2 µm Bio-Inert® membrane, natural 10/pack 5046 3.0 µm glass fiber media/0.2 µm Bio-Inert membrane, natural 10/pack 5031 1.0 µm glass fiber, natural † 10/pack 5031L 1.0 µm glass fiber, natural †, with lid 10/pack 5032 1.0 µm glass fiber, white † 10/pack 5029 0.45 µm Supor® membrane, natural 10/pack 5039 1.2 µm Supor membrane, natural 10/pack 5039L 1.2 µm Supor membrane, natural, with lid 10/pack 5041 Prefilter material/1.2 µm Supor membrane, natural 10/pack 5047 Mustang™ Q ion exchange membrane, natural 5048 7267 Mustang S ion exchange membrane, natural 30-40 µm screen, natural † Glass Fiber plates have polypropylene support †† PTFE plates have polyester support Vacuum Manifold Part Number Description Pkg 5017 Multi-well plate vacuum manifold 1/pkg 10/pack 5014 1 mL receiver plate spacer block 1/pkg 10/pack 5015 350 µL receiver plate spacer block 1/pkg 10/pack 5016 Replacement accessory kit (includes O-ring, gasket and allen wrench) 1/pkg Pkg AcroPrep 96 Filter Plates, 1 mL well Accessories Part Number Description Pkg Part Number Description 5066 3K Omega membrane, natural 5/pack 5225 Adapter collar for centrifugation 2/pkg 5052 0.2 µm GHP membrane, natural 5/pack 5230 Cap mat for incubation 5/pkg 5054 0.45 µm GHP membrane, natural 5/pack 5231 Multi-well plate lids 10/pkg 5055 0.2 µm PTFE membrane, natural †† 5/pack 5056 0.45 µm PTFE membrane, natural †† 5/pack 5051 1.0 µm glass fiber media, natural † 5/pack 5053 3.0 µm glass fiber media/0.2 µm Bio-Inert membrane, natural 5/pack 5065 1.2 µm Supor membrane, natural, with valve mats 5/pack 5062 Mustang Q ion exchange membrane, natural 5/pack 5063 Mustang S ion exchange membrane, natural 5/pack Application by Membrane Type GHP Membrane • Cleavage Reactions • Receptor:Ligand Screens • Bead- and Resin-based Applications • Plasmid Lysate Clearing • HPLC Sample Prep and Clarification • Metabolic Studies • Applications that Use Harsh Organics • Combi-chem Library Screens • General Sample Prep PTFE Membrane • Molecular Synthesis • Drug Synthesis Reactions • Bead- and resin-based Applications Bio-Inert® Membrane • Drug Binding Studies • Purification of Oligos from Polyacrylamide Gels • General Sample Prep Glass Fiber over Bio-Inert Membrane • Clarification of Cell Lysates and Tissue Homogenates • Preparation of Proteinaceous Solutions • Prefiltration Glass Fiber Membrane • Gross Fractionation • Lysate Clarification • Cell-based Assays • DNA Binding Omega™ Ultrafiltration Membrane • Size Exclusion • Concentrate, Purify, and Desalt Proteins, Peptides, Oligos, DNA, and RNA • DNA Clean-up • Recovery of Proteins, Oligos, and RNA from Polyacrylamide Gels Supor® Membrane • Sample Preparation • Plasmid Lysate Purification • Bead- and Resin-based Assays • Receptor:Ligand Binding Assays Related Products Available from Pall • Centrifugal devices for nucleic acid and protein sample preparation in the following sample volumes: Device Sample Volume Nanosep® Device up to 0.50 mL 0.5 to 3.5 mL Microsep™ Device 3 to 15 mL Macrosep® Device 15 to 60 mL Jumbosep™ Device • Nanosep MF (microfiltration) Centrifugal Devices are available with Bio-Inert or GHP membranes for low protein binding and high recoveries in applications such as particulate removal prior to sample analysis (GHP product is HPLC grade) and removal of precipitates. • BioTrace™, Biodyne,® and FluoroTrans® Transfer Membranes offer precise performance and compatibility with nearly every detection system available. • AcroWell™ 96 Filter Plates are an excellent platform for a wide variety of molecular biology detection applications. • Pall Chromatography Resins for the purification of biomolecules and compounds. • Enchant™ Protein Purification Kits expedite proteomic sample prep. References K.J. Valenzano, W. Miller, J. Kravitz, P. Samama, D. Fitzpatrick and K. Seeley. 2000. Development of a Fluorescent Ligand-Binding Assay Using the AcroWell Filter Plate, J. Biomolecular Screening, 5(6):455-461, PN 33220. N. Tang, 2002. Chemiluminescent Detection of Protein Binding, Genetic Engineering News, 22(8):34. Complementary Literature • Product Data, AcroWell 96 Membrane-bottom Filter Plates, PN 33306 • Technical Report, Automated Purification of Combinatorial Libraries Using AcroPrep™ 96 Filter Plate with GHP Membrane, PN 33245 • Protocol, Desalting/Buffer Exchange for Biomolecules Using the AcroPrep 96 UF Filter Plate, PN 33309 • Protocol, Lysate Clearance for Prokaryotic DNA Isolation Using the AcroPrep 96 Filter Plate, PN 33308 • Product Brochure, Accelerate Protein Sample Preparation and Analysis, PN 33411 • Sales Sheet, AcroPrep and AcroWell Multi-well Filter Plates PN 33287 • Scientific Technical Report, The AcroWell Filter Plate Minimizes Crosstalk, online • Protein Sample Preparation and Analysis Application Manual, PN 33465 www.pall.com/lab Pall Life Sciences 600 South Wagner Road Ann Arbor, MI 48103-9019 USA 800.521.1520 (+)800.PALL.LIFE 734.665.0651 734.913.6114 (USA and Canada) (Outside USA and Canada) phone fax Australia – Cheltenham, VIC Tel: 03 9584 8100 1800 635 082 (in Australia) Fax: 1800 228 825 Austria – Wien Tel: 00 1 49 192 0 Fax: 00 1 49 192 400 Canada – Ontario Tel: 905-542-0330 800-263-5910 (in Canada) Fax: 905-542-0331 Canada – Québec Tel: 514-332-7255 800-435-6268 (in Canada) Fax: 514-332-0996 800-808-6268 (in Canada) China – P. 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Pall, , AcroWell, AcroPrep, Biodyne, Bio-Inert, BioTrace, Emflon, Enchant, FluoroTrans, Jumbosep, Macrosep, Microsep, Nanosep, Omega, and Supor are trademarks of Pall Corporation. ® indicates a trademark registered in the USA. is a service mark of Pall Corporation. 8/07, pdf, GN07.1641 PN33296