Automating Electrophoresis With the New Experion™ System

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Applications Tips and Techniques New Products BioRadiations A Resource for Life Science Research Number 115 Cover Story Automating Electrophoresis With the New…

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Applications Tips and Techniques New Products BioRadiations A Resource for Life Science Research Number 115 Cover Story Automating Electrophoresis With the New Experion™ System Features Getting Started in Expression Proteomics Large-Scale Viral Production Embryonic Stem Cell Transfection What’s New High-Fidelity DNA Polymerase More Reaction Vessels and Sealers Chromo4™ Real-Time PCR System Even More Bio-Plex™ Assays And More … AMPLIFICATION REAGENTS A Sea of Amplification Options A complete selection of amplification reagents for both real-time and conventional PCR. ■ Enzymes and kits for a broad array of applications — hot-start PCR, reverse transcription, and real-time PCR using SYBR Green or probe-based detection ■ High-quality reagents designed for convenience — easy-to-use kits and protocols simplify reaction setup to generate consistent, reproducible results ■ Products developed and supported by amplification experts — formulated to provide sensitive detection and linear data over a wide dynamic range To learn more and enter a drawing to win $500* in amplification reagents, visit us on the Web at www.bio-rad.com/ad/ampreagents/ * Open to legal US residents only. For a complete listing of sweepstakes rules, visit www.bio-rad.com/ad/ampreagents/ Practice of the patented polymerase chain reaction (PCR) process requires a license. SYBR is a trademark of Molecular Probes, Inc. Bio-Rad Laboratories, Inc. is licensed by Molecular Probes, Inc. to sell reagents containing SYBR Green I for use in real-time PCR for research purposes only. Visit us on the Web at discover.bio-rad.com Call toll free at 1-800-4BIORAD (1-800-424-6723); outside the US, contact your local sales office. BioRadiations magazine is published by in this issue Bio-Rad Laboratories, Inc. 2000 Alfred Nobel Drive Hercules, CA 94547 USA © 2005 Bio-Rad Laboratories, Inc. Copyright reverts to individual authors upon publication. Reprographic copying for personal BioRadiations 115, 2005 use is allowed, provided credit is given to Bio-Rad Laboratories. Bio-Rad Subsidiary to our readers Telephone Numbers Australia 61-2-9914-2800 Scientific applications of microfluidic technology are rapidly expanding, and have grown to Austria 43-1-877-89-01 include such traditional, routine applications as electrophoresis. Gel electrophoresis is used daily in laboratories worldwide for basic separations and analyses, including determination of sample Belgium 32-9-385-55-11 composition and purity. Recent automation of the electrophoretic process through application of Brazil 55-21-2527-3454 microfluidics now allows rapid, sensitive, unattended runs, complete with basic sample Canada 00-1-905-712-2771 information. Bio-Rad’s long history of developing products for electrophoresis now includes its China 86-21-6426-0808 introduction of the Experion™ automated electrophoresis system. Some of the product Czech Republic 420-2-41-43-05-32 developers describe this exciting new system in this issue’s cover story. Denmark 45-44-52-10-00 Finland 358-9-804-22-00 cover story France 33-1-47-95-69-65 16 The Experion System: Bio-Rad Applies Microfluidics to Automate Germany 49-89-318-84-0 Gel Electrophoresis Greece 30-210-777-4396 B Chang, E Larson, and C Whitman-Guliaev, Bio-Rad Laboratories, Inc., Hercules, CA USA Hong Kong 852-2-789-3300 Hungary 36-1-455-8800 India 91-124-239-8112 dimensions Israel 972-3-951-4127 12 Key Considerations When Starting Comparative Proteomics Italy 39-2-21609-1 A Posch and MG Brubacher, Bio-Rad Laboratories, Inc., Hercules, CA USA Japan 81-3-5811-6270 Korea 82-2-3473-4460 departments Latin America 00-1-305-894-5950 Mexico 55-52-00-05-20 2 What’s New The Netherlands 31-318-540666 32 Tips and Techniques New Zealand 64-9-415-2280 32 New Literature Norway 47-23-38-41-30 Poland 48-22-331-99-99 technical reports Portugal 351-21-472-7700 Russia 7-095-721-1404 22 Feasibility of Multiplexing Bio-Plex™ Total Target and Phosphoprotein Assays Singapore 65-6415-3188 Q Gao, J Blas, and E Gerasimopoulos, Bio-Rad Laboratories, Inc., Hercules, CA USA South Africa 27-11-4428508 24 Cytokine Comparison of Three Tissues Following Genital Herpes Simplex Spain 34-91-590-5200 Virus Type 2 Infection Using the Bio-Plex Suspension Array System and Sweden 46-8-555-12700 Sandwich ELISA Switzerland 41-61-717-9555 DJJ Carr, The Dean A McGee Eye Institute, Oklahoma City, OK USA Taiwan 88-62-2578-7189 27 Highly Efficient Transfection of Mouse ES Cells With TransFectin™ Lipid Reagent Thailand 662-651-8311 S De-Zolt, C Strolz, and J Altschmied, University Hospital Frankfurt, Frankfurt, Germany United Kingdom 44-20-8328-2000 29 Purification of Recombinant Adeno-Associated Virus Type 2 Using USA Toll free 1-800-4BIORAD (1-800-424-6723) UNOsphere™ Media J Zhen, Bio-Rad Laboratories, Inc., Hercules CA USA discover.bio-rad.com Legal Notices On the cover: ABI PRISM is a trademark of Applera Corp. Cibacron is a trademark of Ciba-Geigy Corp. Coomassie is a trademark of BASF The Experion electrophoresis Aktiengesellschaft. Depo-Provera is a trademark of Pfizer Inc. Hoechst is a trademark of Hoechst Aktiengesellschaft. LabChip and the station, part of the Experion automated LabChip logo are registered trademarks of Caliper Life Sciences, Inc. Bio-Rad Laboratories, Inc. is licensed by Caliper Life Sciences, electrophoresis system Inc. to sell products using the LabChip technology for research use only. These products are licensed under US Patent Nos. 5,863,753, 5,658,751, 5,436,134 and 5,582,977, and pending patent applications, and related foreign patents, for internal research and development use only in detecting, quantitating and sizing macromolecules, in combination with microfluidics. Quantikine is a trademark of Research & Diagnostic Systems, Inc. Sepharose is a trademark of Amersham Pharmacia Biotech AB Corp. siGLO is a trademark of Dharmacon, Inc. StrepTactin is a trademark of Institut für Bioanalytik GmbH. StrepTactin is covered by German Patent application P 19641876.3. Bio-Rad Laboratories, Inc. is licensed by Institut für Bioanalytik GmbH to sell this product for research use only. SYPRO is a trademark of Molecular Probes, Inc. Tween is a trademark of ICI Americas Inc. xMAP is a trademark of Luminex Corp. The Bio-Plex suspension array system includes fluorescently labeled microspheres and instrumentation licensed to Bio-Rad Laboratories, Inc. by the Luminex Corp. Practice of the patented polymerase chain reaction (PCR) process requires a license. Bio-Rad and MJ brand thermal cyclers and systems include an Authorized Thermal Cycler and may be used with PCR licenses available from Applied Biosystems. Their use with Authorized Reagents also provides a limited PCR license in accordance with the label rights accompanying such reagents. Some applications may also require licenses from other third parties. BioRadiations 115 1 what’s new Experion™ Automated Electrophoresis System Bio-Rad combines decades of experience in electrophoresis with 2 the innovation of Caliper Life Sciences’ LabChip technology to introduce the Experion automated electrophoresis system, a powerful new 1 system that delivers affordable, accessible automation for protein and RNA electrophoresis to any research laboratory. Rapid, Automated Results The Experion system applies 3 microfluidic separation technology to speed up data acquisition without compromising data quality. It automatically performs all the steps of 8 gel-based electrophoresis (sample separation, staining, 4 destaining, imaging, band 5 6 7 Experion system components 1. Electrophoresis station detection, and even some data analysis) to 2. Data analysis software deliver reproducible separation and quantitation results in just 30 minutes. 3. Vortex station 4. Analysis kit reagents for protein Integrated System Design and Superior Performance or RNA samples 5. RNA HighSens analysis chip Sleek, user-friendly components exemplify the quality and innovation of the 6. RNA StdSens analysis chip Experion system. Microfluidic chips combine with electrophoresis-grade 7. Pro260 analysis chip reagents, automated electrophoresis and priming stations, an easy-to-use vortex 8. Priming station station, and powerful analysis software to form an integrated system that streamlines the analytical process. ã Fast, 30 minute batch runs of protein and RNA samples ã Accurate single-step protein sizing from 10 to 260 kD ã Protein resolution and sensitivity comparable to colloidal Coomassie Blue-stained mini gels ã 2-in-1 process for RNA: integrity checks and quantitation ã Optimized reagents and exclusive protein and RNA standards produce accurate, reproducible sizing and quantitation ã Flexible software tools make data analysis easy and efficient For more information, request bulletin 3140. Automation Reduces Hands-On Time Required for Electrophoresis The Experion system includes three instruments that automate and expedite the electrophoresis process: ã The electrophoresis station combines the steps of gel-based electrophoresis into a single walk-away process ã The priming station prepares microfluidic chips for electrophoresis — Preset time and pressure settings ensure optimal, consistent introduction of gel solution into the microchannels of the chips, in preparation for sample separation and analysis ã The vortex station ensures sufficient mixing of RNA analysis reagents for effective sample runs All three instruments have been optimized to minimize user intervention. High-Quality Chips and Reagents Ensure Exceptional Resolution and Quantitation Experion analysis kits combine innovative chip design with high-quality reagents to perform reproducible, quantitative, and accurate protein and RNA analyses in minutes. Streamlined chip preparation methods and minimal sample requirements result in rapid experiments with minimal hands-on time. Kits are available in flexible ordering configurations to match your research needs. 2 BioRadiations 115 © 2005 Bio-Rad Laboratories, Inc. what’s new Each analysis kit includes the following: 30 mm ã Experion Pro260, RNA StdSens, or RNA HighSens chips ã Optimized gel solution for separation and resolution comparable to results achieved with mini gels ã High-sensitivity fluorescent dye for optimal detection ã Protein or RNA ladder for accuracy in sizing and quantitation ã Sample buffer for optimized, reproducible running conditions Specifications for Experion Analysis Kits 50 mm Pro260 Chip RNA StdSens Chip RNA HighSens Chip Reagents RNA Ladder Number of samples 1–10 1–12 1–11 — — Sample volume 4 μl 1 μl 1 μl — — Dynamic range 2.5–2,000 ng/μl 5–500 ng/μl 100–5,000 pg/μl — — Separation range 10–260 kD 100–6,000 bases 100–6,000 bases — — Sensitivity 2.5 ng/μl of carbonic 5 ng 100 pg — — anhydrase in 1x PBS Shelf life 9 months 9 months 9 months 9 months 9 months Storage conditions Ambient Ambient Ambient 4ºC –20ºC Caddy dimensions (W x D x H) 30 x 10 x 50 mm 30 x 10 x 50 mm 30 x 10 x 50 mm — — For more information, request bulletins 3169 and 3170. Flexible Software Tools for Efficient Data Analysis Experion software simplifies your workflow with convenient icons and pull-down menus, and convenient data analysis tools allow you to quickly obtain the information you need. The screen displays separation results in both electropherogram (peak) and simulated gel views. The software automatically calculates protein or RNA component size, concentration, and percent of total sample. Results for each peak appear in the Results table. ã Sizing and quantitation calculations performed automatically ã Intuitive navigation of separation and data analysis screens ã Quick comparisons of protein or RNA components across all samples in the chip ã Multiple printing and data export options For more information, request bulletin 3171. For a complete literature packet on the Experion system, request bulletin 3174A. Ordering Information Catalog # Description Experion LabChip Technology. Experion Automated Electrophoresis System The Experion analysis chip 700-7000 Experion System, 100–120/220–240 V, for protein analysis, includes electrophoresis station, priming station, houses LabChip technology software, USB2 cable, test chip, instructions (analysis kits sold separately) developed by Caliper Life 700-7001 Experion System, 100–120 V, for RNA analysis, includes electrophoresis station, priming station, vortex station, Sciences, Inc. The chip is a software, USB2 cable, test chip, instructions (analysis kits sold separately) powerful, miniaturized device 700-7002 Experion System, 220–240 V, for RNA analysis, includes electrophoresis station, priming station, vortex station, (much like a tiny laboratory) software, USB2 cable, test chip, instructions (analysis kits sold separately) that combines the functionality Experion Analysis Kits of several larger benchtop 700-7101 Experion Pro260 Analysis Kit for 10 Chips, includes 10 Pro260 chips, Experion Pro260 reagents analytical instruments. Up to and supplies for 10 chips 10 protein or 12 RNA samples 700-7102 Experion Pro260 Analysis Kit for 25 Chips, includes 25 Pro260 chips, Experion Pro260 reagents can be analyzed in only 30 and supplies for 25 chips minutes. (Chip shown at 700-7103 Experion RNA StdSens Analysis Kit for 10 Chips, includes 10 RNA StdSens chips, Experion approximately actual size.) RNA StdSens reagents and supplies for 10 chips 700-7104 Experion RNA StdSens Analysis Kit for 25 Chips, includes 25 RNA StdSens chips, Experion RNA StdSens reagents and supplies for 25 chips 700-7105 Experion RNA HighSens Analysis Kit for 10 Chips, includes 10 RNA HighSens chips, Experion RNA HighSens reagents and supplies for 10 chips 700-7106 Experion RNA HighSens Analysis Kit for 25 Chips, includes 25 RNA HighSens chips, Experion RNA HighSens reagents and supplies for 25 chips Visit us on the Web at discover.bio-rad.com BioRadiations 115 3 what’s new Model 680 Microplate Reader Updated The popular Model 680 has been updated to make it the most flexible and versatile microplate reader in the marketplace today. This microplate reader has always been an excellent choice for visible-light colorimetry, with its onboard software, plate-shaking function, 8-position filter wheel, and versatility to read both U- and V-bottom plates and 8- and 12-well strips. The updated version offers even more with these features: ã Clearer LCD display — now 5 mm thicker ã Choice of integrated English and Chinese firmware ã Onboard storage of standard curves and graphs ã Ability to print graphics on external printer — ASCII or ESCIP type formats ã Self-diagnostic capabilities to detect lamp burnout at startup Ordering Information Catalog # Description 168-1000 Model 680 Microplate Reader, 100/240 V, 50/60 Hz, includes 415, 450, 490, and 655 nm filters (installed), power cord, spare fuse, dust cover, instructions 168-1001 Model 680 Microplate Reader With Temperature Control, 100/240 V, 50/60 Hz, includes 415, 450, 490, and 655 nm filters (installed), power cord, spare fuse, dust cover, instructions 168-1002 Model 680 Microplate Reader With Internal Thermal Printer, 100/240 V, 50/60 Hz, includes 415, 450, 490, and 655 nm filters (installed), 1 roll of printer paper, power cord, spare fuse, dust cover, instructions 168-1003 Model 680 Microplate Reader With Temperature Control and Internal Thermal Printer, 100/240 V, 50/60 Hz, includes 415, 450, 490, and 655 nm filters (installed), 1 roll of printer paper, power cord, spare fuse, dust cover, instructions 168-1100 Model 680 XR Microplate Reader, 100/240 V, 50/60 Hz, includes 340, 405, 415, and 655 nm filters (installed), power cord, spare fuse, dust cover, instructions 168-1101 Model 680 XR Microplate Reader With Temperature Control and Internal Thermal Printer, 100/240 V, 50/60 Hz, includes 340, 405, 415, and 655 nm filters (installed), 1 roll of printer paper, power cord, spare fuse, dust cover, instructions EXQuest™ Spot Cutter Rapid Excision With Professional Performance The EXQuest spot cutter is a next-generation gel excision robot from Bio-Rad. The design includes functionality and value for every laboratory, from occasional precision cutting to high-throughput, multi-gel imaging and cutting. The EXQuest spot cutter is desirable for any laboratory where precision gel cutting and spot identification are required. Benefits include: ã Cutting of over 600 spots/hr ã Illumination for white-light and UV applications ã Cutting of gels and blots ã Full automation of 2-D and 1-D gel cutting ã Walk-away performance ã Precision cutting with 100 µm resolution ã Liquid-handling capabilities for accurate and reproducible The EXQuest spot cutter is fully automated, with a removable gel support performance tray and four-microplate capacity. ã Hands-free processing of up to 4 precast gels ã Streamlined workflow with PDQuest™ 2-D and Ordering Information Quantity One® 1-D analysis software Catalog # Description 165-7200 EXQuest Spot Cutter 165-7201 EXQuest Spot Cutter With PC 4 BioRadiations 115 © 2005 Bio-Rad Laboratories, Inc. what’s new Protein Blotting Starter Kits The color blot and chemi blot protein blotting starter kits contain all the tools you need for western blotting experiments. The kits contain all materials and reagents required to: ã Transfer proteins to PVDF and nitrocellulose membranes with Tris/glycine transfer buffer ã Block nonspecific sites with nonfat milk ã Wash the membrane and reduce nonspecific binding with TBS and 10% Tween 20 ã Label the protein of interest with goat anti-rabbit or goat anti- mouse secondary antibody conjugated to horseradish peroxidase (HRP) or alkaline phosphatase (AP) ã Detect the protein with your choice of AP- or HRP-mediated chemiluminescent or colorimetric substrate ã Accurately determine molecular weight from blots using unstained or prestained Precision Plus Protein™ standards The protein blotting guide, included with each protein blotting starter kit, will guide you through western blots from Protein Blotting Guide start to finish. This booklet provides information on protein transfer and detection chemistries, methods, equipment, and reagents used in protein blotting. The guide also offers helpful troubleshooting and technical advice. For additional copies of the protein blotting guide, request bulletin 2895. A Guide to Transfer and Detection Third Edition For more information, request bulletin 3117. Ordering Information Catalog # Description 170-5050 HRP Color Blot Starter Kit 1, includes 1 L 4CN (HRP substrate), 2 sheets Immun-Blot® PVDF membrane, 2 sheets 0.2 μm nitrocellulose membrane, 50 μl Precision Plus Protein™ Kaleidoscope™ standards, 1 L 10x TBS, 100 ml 10% Tween 20, 300 g nonfat dry milk, 2 ml GAR-HRP conjugate, 10x Tris/glycine buffer 170-5051 HRP Color Blot Starter Kit 2, same as 170- 5050 except with 2 ml GAM-HRP conjugate 170-5052 AP Color Blot Starter Kit 1, includes 1 L BCIP/NBT (AP substrate), 2 sheets Immun- Blot PVDF membrane, 2 sheets 0.2 μm nitrocellulose membrane, 50 μl Precision Plus Protein Kaleidoscope standards, 1 L 10x TBS, 100 ml 10% Tween 20, 300 g nonfat dry milk, 2 ml GAR-AP conjugate, 10x Tris/glycine buffer 170-5053 AP Color Blot Starter Kit 2, same as 170-5052 except with 2 ml GAM-AP conjugate 170-5054 HRP Chemi Blot Starter Kit 1, includes 100 ml Immun-Star HRP substrate, 2 sheets Immun- Blot PVDF membrane, 2 sheets 0.2 μm nitrocellulose membrane, 50 μl Precision Plus Protein unstained standards, 0.3 ml Precision Protein StrepTactin-HRP conjugate, 1 L 10x TBS, 100 ml 10% Tween 20, 300 g nonfat dry milk, 2 ml GAR-HRP conjugate, 10x Tris/glycine buffer 170-5055 HRP Chemi Blot Starter Kit 2, same as 170- 5054 except with 2 ml GAM-HRP conjugate 170-5056 AP Chemi Blot Starter Kit 1, includes 100 ml Immun-Star AP substrate, 2 sheets Immun-Blot PVDF membrane, 2 sheets 0.2 μm nitrocellulose membrane, 50 μl Precision Plus Guide to protein blotting starter kits. Choose an appropriate detection method. The catalog numbers Protein unstained standard
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