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Summary of Contents for BIO RAD Bio-Scale
- Page 1 Bio-Scale Mini ™ Profinity eXact ™ Cartridges, 1 and 5 ml Instruction Manual Catalog # 732-4646 732-4648...
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Page 2: Table Of Contents
Table of Contents Section 1 ...Introduction ........1 Section 2 Product Information......2 Section 3 Connection to Low-Pressure Chromatography Systems ....7 Bio-Rad Laboratories, Inc. Section 4 Connection to Medium- and High- 2000 Alfred Nobel Dr. Pressure Chromatography Systems .11 Hercules, CA 94547 USA BioLogic DuoFlow ™... -
Page 3: Section 1
Section 14 Legal Notices ........29 exchange, hydrophobic interaction, and affinity chromatography. Bio-Scale Mini Profinity eXact cartridges are packed with Profinity eXact (EXact Affinity Cleavage Technology) purification resin. This agarose-based affinity chromatography resin utilizes an immobilized, modified... -
Page 4: Section 2 Product Information
Bio-Scale Mini cartridges are prepacked inner diameter chromatography cartridges supplied ready for use in 5 ml: 40 mm length x 12.6 mm 1 ml and 5 ml sizes. Bio-Scale Mini cartridges can inner diameter Maximum pressure tolerance 45 psi be used with any liquid chromatography system... - Page 5 Do not use. Protease PMSF, CalBioChem cocktail, inhibitors Roche Protease Inhibitor cOmplete mini tablet BD Biosciences cocktail Lysis Solutions Lysis & Extractrion Reagent (Bio-Rad), B-PER phosphate (Pierce), BugBuster (EMD), FastBreak Cell Lysis (Promega) ReadyPreps Lysis (Epicentre), Do not use. CelLytic Express (Sigma)
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Page 6: Section 3 Connection To Low-Pressure Chromatography Systems
Chromatography Systems At 8 M, binding capacity and target protein purity will be reduced. Bio-Scale Mini cartridges are ideal for use with Other additives CaCl £5 mM when used with Bio-Rad's BioLogic™ LP chromatography system, MES, MOPS, or PIPES buffers. - Page 7 1.6 mm ID tubing, then connect to the top of (if available). If using the MV-6 sample inject the female luer on the Bio-Scale Mini cartridge. valve, turn the knob counterclockwise as far as For optimum performance, a cartridge should...
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Page 8: Pressure Chromatography Systems
1.6 mm ID tubing, then connect to the bottom liquid chromatography system, provided that the of the male luer on the Bio-Scale Mini cartridge. system does not exceed the maximum pressure limit of the cartridges (3 bar, 45 psi or 300 kPa). It is recommended that the system pressure limit be set according to the cartridge pressure limit. -
Page 9: Hplc Systems
The luer to 10-32 adaptor fittings kit (catalog rate of the cartridge. #732-0112) provides fittings necessary to connect the Bio-Scale Mini cartridge to nut and ferrule type * Fittings kit ordering information can be found within fittings found on most HPLC systems. Alternatively, the Ordering Information section of this manual. -
Page 10: Section 5
100 mM sodium phosphate, pH 7.2 Buffers and Methods Elution buffer* 100 mM sodium phosphate, 100 mM sodium fluoride, pH 7.2 Bio-Scale Mini Profinity eXact cartridges can be run Regeneration 100 mM phosphoric acid Storage 0.02% sodium azide, using either native or denaturing purification protocols. -
Page 11: Section 6 Quick Solubility Screening Protocol
Section 6 supernatant to a clean tube. Remove 50 µl of the supernatant, and label the tube "Soluble". Quick Solubility Screening Resuspend the insoluble pellet in 500 µl of Protocol bind/wash buffer containing 4 M urea. Centrifuge the lysate at 16,000 x g for 5 min at Before choosing a native or denaturing purification 4°C. -
Page 12: Section 7 Preparation Of E. Coli Lysate
Section 7 Preparation of E. coli Lysate For E. coli cultures expressing medium to high levels of ≥ Profinity eXact-tagged proteins, ( 10% of total protein), 50 ml of culture will yield sufficient material for a 1 ml cartridge purification, and 250 ml of culture will yield sufficient material for a 5 ml cartridge purification run. -
Page 13: Section 8 Cartridge Preparation And Purification Protocol
Remove the supernatant, and filter through a Purification Protocol 0.45 µM filter before applying to the cartridge. Equilibrate the cartridge with 10 column volumes Section 8 (CV) of bind/wash buffer at 3 ml/min (10 ml/min). Load the sample lysate at 1 ml/min (5 ml/min). Cartridge Preparation and Wash the cartridge with 10 CV of bind/wash Purification Protocol... -
Page 14: Section 9 Scaling Up
Tube #: 1 2 3 4 567 1112 Scaling Up 2.00 1.75 Bio-Scale Mini cartridges are available in 1 ml and 1.50 1.25 5 ml cartridge formats. The Profinity eXact resin is 1.00 also available in 10 ml bottles. For quick scale-up, 0.75... - Page 15 cartridge after five purifications, however, the following Note: The resin may also be cleaned with regeneration procedure may be used to prolong the 0.1 M NaOH. Limit exposure time to 3 hours useful lifespan of a cartridge. In addition, in order to and immediately neutralize with 5 CV of avoid cross-contamination it is recommended that bind/wash buffer.
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Page 16: Section 11 Troubleshooting Guide
Section 11 Problem Possible Cause Solution Precipation during Binding capacity of Load less sample Troubleshooting Guide purification cartridge exceeded Protein aggregation Include detergent or glycerol in buffers Problem Possible Cause Solution Cartridge clogging Particulates in samples Filter all samples and buffers or slow flow rate or buffers through 0.45 µM filter prior... -
Page 17: Section 12 Ordering Information
Cartridges triggered processing protease useful for one-step Catalog # Description purification. Biochemistry 43, 14539-46 (2004) 732-4646 Bio-Scale Mini Profinity eXact cartridge, 2 x 1 ml cartridge Section 14 732-4648 Bio-Scale Mini Profinity eXact cartridge, 1 x 5 ml cartridge Legal Notices For the most up to date list of other cartridge offerings, please visit online at www.bio-rad.com/cartridges/... - Page 18 Affinity Proteins. This product is intended for research purposes only. For commercial applications or manufacturing using these products, commercial licenses can be obtained by contacting the Life Science Group Chromatography Marketing Manager, Bio-Rad Laboratories, Inc., 6000 Alfred Nobel Drive, Hercules, CA 94547, Tel (800) 4BIORAD.