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Bio-Scale

™

Mini

Profinity eXact

™

Cartridges, 1 and 5 ml

Instruction Manual

Catalog #
732-4646
732-4648

10011164A.qxp 11/19/2007 9:54 AM Page I

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Содержание Bio-Scale

Страница 1: ...Bio Scale Mini Profinity eXact Cartridges 1 and 5 ml Instruction Manual Catalog 732 4646 732 4648 10011164A qxp 11 19 2007 9 54 AM Page I...

Страница 2: ...1 4 1 BioLogic DuoFlow Systems 11 4 2 HPLC Systems 12 4 3 FPLC Systems 13 Section 5 Buffers and Methods 14 Section 6 Quick Solubility Screening Protocol 16 Section 7 Preparation of E coli Lysate 19 Se...

Страница 3: ...omatography resin utilizes an immobilized modified protease that selectively binds the fusion protein and cleaves the affinity tag on column under controlled conditions releasing the purified target p...

Страница 4: ...Storage recommendations 100 mM soidum phosphate 0 02 sodium azide pH 7 2 Autoclavability Not autoclavable 3 Section 2 Product Information Bio Scale Mini cartridges are prepacked chromatography cartrid...

Страница 5: ...tease PMSF CalBioChem cocktail inhibitors Roche Protease Inhibitor 1X cOmplete mini tablet BD Biosciences cocktail 2X Lysis Solutions Lysis Extractrion Reagent 1X Bio Rad B PER phosphate Pierce BugBus...

Страница 6: ...s it will go then turn clockwise three full turns Assemble with fittings and lock rings as shown in Figure 1 US patent 5 135 658 7 Reagent Group Description Stability Denaturants Guanidine HCl Do not...

Страница 7: ...STEM OR ECONO GRADIENT PUMP MV 6 INJECT VALVE SAMPLE LOOP WASTE INJECT PORT F IL L TO COLUMN MV 6 INJECT VALVE SAMPLE LOOP WASTE INJECT PORT FILL INJECT POSITION FILL POSITION 9 Fig 1 BioLogic LP syst...

Страница 8: ...ype system 4 1 BioLogic DuoFlow Systems The luer to BioLogic system fittings kit catalog 732 0113 includes 1 4 28 female to male luer and 1 4 28 female to female luer to connect one Bio Scale Mini car...

Страница 9: ...ommended flow rate of the cartridge Fittings kit ordering information can be found within the Ordering Information section of this manual 13 Fig 4 Luer to 1 4 28 adaptor 4 2 HPLC Systems The luer to 1...

Страница 10: ...cale Mini Profinity eXact cartridges can be run using either native or denaturing purification protocols Under native conditions proteins are purified using buffers that help retain the natural folded...

Страница 11: ...creening Protocol Before choosing a native or denaturing purification protocol it is useful to determine the approximate expression level of a protein and to determine if the overexpressed target prot...

Страница 12: ...cell pellet and resuspend in 10 volumes of bind wash buffer 50 ml of culture typically yields 0 5 g of paste resulting in approximately 5 ml of lysate 3 Sonicate the lysate on ice for 3 minutes 4 Cent...

Страница 13: ...may be necessary for complete cleavage of some proteins 21 5 Remove the supernatant and filter through a 0 45 M filter before applying to the cartridge Section 8 Cartridge Preparation and Purificatio...

Страница 14: ...ge overuse It is recommended to dispose of a 23 Fig 6 Native purification Purification of Profinity eXact tagged Maltose Binding Protein from the soluble fraction using a BioLogic DuoFlow system 5 ml...

Страница 15: ...d for single proteins In order to reuse a cartridge the Profinity eXact tag which is bound tightly to the functional ligand MUST be removed Therefore to maintain good cartridge flow properties and to...

Страница 16: ...lysate to degrade DNA No target protein Low level of target Check expression level by in eluant protein in starting SDS PAGE material Target protein not Used resin must be binding regenerated before r...

Страница 17: ...ription 732 4646 Bio Scale Mini Profinity eXact cartridge 2 x 1 ml cartridge 732 4648 Bio Scale Mini Profinity eXact cartridge 1 x 5 ml cartridge For the most up to date list of other cartridge offeri...

Страница 18: ...tended 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 Ma...

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