Bio-rad Profinity eXact™ Purification and Tag Cleavage Con Manual de usuario Pagina 1

Bio-Scale™Mini Profinity eXact™Cartridges, 1 and 5 mlInstruction ManualCatalog #732-4646732-464810011164A.qxp 11/19/2007 9:54 AM Page I

Table 4. Buffer composition.Bind/wash buffer* 100 mM sodium phosphate, pH 7.2Elution buffer* 100 mM sodium phosphate,100 mM sodium fluoride, pH 7.2Reg

supernatant to a clean tube. Remove 50 µl ofthe supernatant, and label the tube "Soluble".3. Resuspend the insoluble pellet in 500 µl ofbin

Section 7Preparation of E. coli LysateFor E. coli cultures expressing medium to high levels ofProfinity eXact-tagged proteins, (≥10% of total protein)

Purification Protocol1. Equilibrate the cartridge with 10 column volumes(CV) of bind/wash buffer at 3 ml/min (10 ml/min).2. Load the sample lysate a

Section 9Scaling UpBio-Scale Mini cartridges are available in 1 ml and 5 ml cartridge formats. The Profinity eXact resin isalso available in 10 ml bot

Note: The resin may also be cleaned with 0.1 M NaOH. Limit exposure time to 3 hoursand immediately neutralize with 5 CV ofbind/wash buffer. Regenerati

Problem Possible Cause SolutionPrecipation during Binding capacity of Load less samplepurification cartridge exceededProtein aggregation Include deter

Section 13ReferencesRuan, B et al. Engineering subtilisin into a fluoride-triggered processing protease useful for one-steppurification. Biochemistry

Profinity eXact™vectors, tags, and resins are exclusively licensed under patent rights of PotomacAffinity Proteins. This product is intended forresear

Table of ContentsSection 1...Introduction ...1Section 2 Product Information...2Section 3

Section 1IntroductionBio-Scale Mini cartridges are convenient, disposable,prepacked chromatography cartridges. Patented*column design assures leak-fre

Table 1. Bio-Scale Mini Profinity eXact cartridge specifications.Sizes 1 ml and 5 ml bed volumesDimensions 1 ml: 40 mm length x 5.6 mm inner diamete

Table 3. Buffer and chemical compatibilities forProfinity eXact cartridges.ReagentGroup Description StabilityTriggering Ions F-, Cl-, N3-, NO2-, CO2-D

Section 3Connection toLow-PressureChromatography SystemsBio-Scale Mini cartridges are ideal for use with Bio-Rad's BioLogic™ LP chromatography sy

Fig. 2. Connecting to an MV-6 valve.3. Connect the inlet of the cartridge to the maleluer fitting on the MV-6 sample inject valve (seeFigure 2). If n

Section 4Connection to Medium and High-PressureChromatography SystemsBio-Scale Mini cartridges can be connected to anyliquid chromatography system, pr

4.3 FPLC SystemsThe luer to M6 adaptor fittings kit (catalog #732-0111)provides fittings necessary to connect the Bio-ScaleMini cartridge to the M6 f