GE HEALTHCARE AKTAprime plus Quick Reference Instructions
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GE HEALTHCARE AKTAprime plus Quick Reference Instructions

GE HEALTHCARE AKTAprime plus Quick Reference Instructions

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ÄKTAprime plus
Cue Cards
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Summary of Contents for GE HEALTHCARE AKTAprime plus

  • Page 1: Table Of Contents

    GE Healthcare ÄKTAprime plus Cue Cards Contents System Preparation Buffer exchange on HiTrap Desalting Buffer exchange on HiPrep 26/10 Desalting Histidine-tagged protein purification, step elution Histidine-tagged protein purification, gradient elution IMAC purification - any metal On-column refolding GST-tagged protein purification...

  • Page 2: System Preparation

    System Preparation Note: Wait at least 15 minutes before using the Starting the system system after the lamp has been turned on to avoid a) Press the button on the back of the system. drifting UV base line. b) Wait until the system self-test has been completed c) If using pH: (30-40 seconds).
  • Page 3 e) Place the tubing holder over the length guide (small c) Connect the column between port 1 on the injection hole) in the delivery arm, push the tubing down to the valve (7-port valve) and the UV flow cell. bottom of the guide and tighten the nut. This ensures that the correct length of the tubing is exposed.

  • Page 4: Buffer Exchange On Hitrap Desalting

    Buffer exchange on HiTrap Desalting c) Enter the sample volume and press to start the Preparing the buffers template. • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. Desalting HiTrap Desalting Theoretical gradient in Application Template.
  • Page 5 Troubleshooting Ordering information High backpressure: Product Quantity Code No. HiTrap Desalting 5 × 5 ml 17-1408-01 • Column clogged – Clean the column according 100 × 5 ml 11-0003-29 to instructions. Make sure the sample has been centrifuged and/or filtered through a 0.45 µm filter. Superloop 10 ml 18-1113-83 •...

  • Page 6: Buffer Exchange On Hiprep 26/10 Desalting

    Buffer exchange on HiPrep 26/10 Desalting c) Enter the sample volume and press to start the Preparing the buffers template. • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. Desalting HiPrep Desalting Theoretical gradient in Application Template.
  • Page 7 Troubleshooting Ordering information High backpressure: Product Quantity Code No. • Column clogged – Clean the column according HiPrep 26/10 Desalting 17-5087-01 (53 ml) to instructions. Make sure the sample has been 17-5087-02 (53 ml) centrifuged and/or filtered through a 0.45 µm filter. Superloop 10 ml 18-1113-83 •...

  • Page 8: Histidine-Tagged Protein Purification, Step Elution

    Histidine-tagged protein purification, step elution e) Connect a sample loop large enough for your sample Preparing the buffers between port 2 and 6 on the injection valve. Use a • Use high purity water and chemicals. syringe to manually fill the loop. •...
  • Page 9 Ordering information Typical result Product Quantity Code No. Sample: Clarified homogenate of E. coli expressing histidine-tagged protein HisTrap HP 5 × 1 ml 17-5247-01 Column: HisTrap™ HP 1 ml 100 × 1 ml 17-5247-05 Binding buffer (port A1): 20 mM phosphate, 0.5 M NaCl, 20 mM imidazole, pH 7.4 HisTrap FF 5 ×...

  • Page 10: Histidine-Tagged Protein Purification, Gradient Elution

    Histidine-tagged protein purification, gradient elution Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 11 Ordering information Typical result Product Quantity Code No. Sample: Clarified homogenate of E. coli expressing histidine-tagged protein HisTrap HP 5 × 1 ml 17-5247-01 Column: HisTrap HP 1 ml Binding buffer (port A1): 20 mM phosphate, 0.5 M NaCl, 20 mM 100 ×...

  • Page 12: Imac Purification - Any Metal

    IMAC purification - any metal Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 13 Ordering information Typical result Product Quantity Code No. Sample: Clarified homogenate of E. coli expressing histidine-tagged protein HiTrap IMAC HP 5 x 1 ml 17-0920-03 Column: HiTrap Chelating HP 1 ml Binding buffer (port A1): 20 mM phosphate, 0.5 M NaCl, HiTrap IMAC FF 5 x 1 ml 17-0921-02...

  • Page 14: On-Column Refolding

    On-column refolding Note: If a Superloop is needed, additional information Preparing the buffers is supplied in the instructions for Superloop. • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. Selecting Application Template and Binding buffer (port A1): starting the method 6 M guanidine hydrochloride, 20 mM Tris-HCl, 0.5 M NaCl,...
  • Page 15 5 Typical result Ordering information Product Quantity Code No. Sample: Clarified homogenate of E. coli expressing histidine-tagged protein HisTrap HP 5 × 1 ml 17-5247-01 Column: HisTrap FF 1 ml Binding buffer (A1): 6 M Guanidine hydrochloride, 20 mM 100 × 1 ml 17-5247-05 Tris-HCl, 0.5 M NaCl, 5 mM Imidazole, 1mM 2-mercaptoethanol, pH 8.0...

  • Page 16: Gst-Tagged Protein Purification

    GST-tagged protein purification Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 17 Ordering information Typical result Product Quantity Code No. Sample: Clarified homogenate of E. coli expressing GST-tagged protein GSTrap FF 2 × 1 ml 17-5130-02 Column: GSTrap™ FF 1 ml Binding buffer (A1): 20 mM sodium phosphate, 0.15 M NaCl, pH 7.3 5 ×...

  • Page 18: Strep(Ii)-Tagged Protein Purification

    Strep(II)-tagged protein purification Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 19 Ordering information Typical result Product Quantity Code No. Sample: Clarified lysate of E. coli expressing Strep(II)-tagged protein. StrepTrap HP 5 × 1 ml 28-9075-46 Column: StrepTrap™ HP 1 ml Binding buffer (port A1): 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, pH 8.0 HiTrap Desalting 5 ×...

  • Page 20: Mbp-Tagged Protein Purification

    MBP-tagged protein purification Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 21 Ordering information Typical result Product Quantity Code No. Sample: Clarified lysate of E. coli expressing MBP-tagged protein. Column: MBPTrap™ HP 1 ml MBPTrap HP 5 × 1 ml 28-9187-78 Binding buffer (port A1): 20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.4 HiTrap Desalting 5 ×...

  • Page 22: Mab Purification, Step Elution

    MAb purification, step elution Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers thtough a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 23 Ordering information Typical result Product Quantity Code No. 280 nm –– UV 280 nm –– HiTrap Protein G HP 2 × 1 ml 17-0404-03 5 × 1 ml 17-0404-01 Sample: Cell culture supernatant HiTrap Protein A HP 2 × 1 ml 17-0402-03 containing mouse IgG 1 Column: HiTrap Protein G HP 1 ml...

  • Page 24: Mab Purification, Gradient Elution

    MAb purification, gradient elution Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1):...
  • Page 25 Ordering information Typical result Product Quantity Code No. Sample: Cell culture supernatant containing mouse IgG 2a HiTrap Protein G HP 2 × 1 ml 17-0404-03 Column: HiTrap Protein A HP 1 ml Binding buffer (A1): 0.1 M sodium phosphate, 0.1 M sodium citrate, pH 7.0 5 ×...

  • Page 26: Igm Purification

    IgM purification Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1): 20 mM sodium phosphate, 0.8 M (NH...
  • Page 27 Ordering information Typical result Product Quantity Code No. Sample: Hybridoma cell culture containing IgM Column: HiTrap IgM Purification HP 1 ml HiTrap IgM Purification HP 5 × 1 ml 17-5110-01 Binding buffer (A1): 20 mM sodium phosphate, 0.8 M (NH 4 ) 2 SO 4 , pH 7.5 Elution buffer 1 (B): 20 mM sodium phosphate, pH 7.5 HiTrap Desalting...

  • Page 28: Albumin Removal

    Albumin removal Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Binding buffer (port A1): 20 mM sodium phosphate, pH 7.0...
  • Page 29 Ordering information Typical result Product Quantity Code No. Sample: Human plasma, buffer exchanged to binding buffer using HiTrap Desalting HiTrap Blue HP 5 × 1 ml 17-0412-01 Column: HiTrap Blue HP 1 ml Binding buffer (A1): 20 mM sodium phosphate, pH 7.0 HiTrap Desalting 5 ×...

  • Page 30: Removal Of Trypsin-Like Serine Proteases

    Removal of trypsin-like serine proteases prime should be displayed. Preparing the buffers b) Use the arrow and OK buttons to move in the menu • Use high purity water and chemicals. tree until you find Affinity Purification any HiTrap. • Filter all buffers through a 0.45 µm filter before use.
  • Page 31 Ordering information Typical result Product Quantity Code No. Sample: Human plasma Column: HiTrap Benzamidine FF (high sub) 1 ml HiTrap Benzamidine FF 2 × 1 ml 17-5143-02 Binding buffer (port A1): 50 mM Tris-HCl, 0.5 M NaCl, pH 7.4 Elution buffer (port B): 50 mM Glycine-HCl, pH 3.0 (high sub) 5 ×...

  • Page 32: Anion Exchange

    Anion exchange Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers through a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Start buffer (port A1): 20 mM Tris-HCl, pH 8.0...
  • Page 33 Ordering information Typical result Product Quantity Code No. Sample: Protein mix containing transferrin, ovalbumin and ß-lactoglobulin in start buffer HiTrap Q HP 5 × 1 ml 17-1153-01 Column: HiTrap Q HP 1 ml Start buffer (A1): 20 mM Tris-HCl, pH 8.0 HiTrap Q FF 5 ×...

  • Page 34: Cation Exchange

    Cation exchange Preparing the buffers Selecting Application Template and starting the method • Use high purity water and chemicals. • Filter all buffers thtough a 0.45 µm filter before use. a) Check the communication to PrimeView. At the lower right corner of the screen the text Controlled By: Start buffer (port A1): 50 mM sodium acetate, pH 5.5...
  • Page 35 Ordering information Typical result Product Quantity Code No. Sample: Protein mix containing chymotrypsinogen A, cytochrome C and lysozyme in start buffer HiTrap SP HP 5 × 1 ml 17-1151-01 Column: HiTrap SP HP 1 ml Start buffer (A1): 20 mM sodium acetate, pH 5.5 HiTrap SP FF 5 ×...

  • Page 36: Method Templates Value Table

    Method Templates value table The Method Templates provides an easy and fast way to This table contains suggested values for a selection of build up a method. By setting defined parameters (and columns which enables a fast start. The values can then volumes) for each part of the chromatographic phase a be optimized to suit the specific application.
  • Page 37 Code No. Column Pressure* Flow Frac.vol Equil.vol Wash1 vol. Elu.vol Wash2 vol. ml ml/min 17-0406-01 HiTrap Heparin HP, 1 ml 17-0407-03 HiTrap Heparin HP, 5 ml 17-5112-01 HiTrap Streptavidin HP, 1 ml 17-5143-01 HiTrap Benzamidine FF (high sub), 1 ml 17-5144-01 HiTrap Benzamidine FF (high sub), 5 ml 17-5189-01 HiPrep 16/10 Heparin FF...
  • Page 38 Code No. Column Pressure* Flow Frac.vol Equil.vol Wash1 vol. Elu.vol Wash2 vol. ml ml/min 17-5092-01 HiPrep 16/10 Q XL 17-5093-01 HiPrep 16/10 SP XL 17-5091-01 HiPrep 16/10 CM FF 17-5090-01 HiPrep 16/10 DEAE FF 17-1064-01 HiLoad™ 16/10 Q Sepharose HP 17-1137-01 HiLoad 16/10 SP Sepharose HP Hydrophobic interaction 17-1355-01 HiTrap Phenyl FF (high sub),...
  • Page 39 11-0027-48 AC, 2007-09 • p39...
  • Page 40 All goods and services are sold subject to the terms and Design, GSTPrep, GSTrap, HisPrep, HisTrap, HiPrep, HiLoad, conditions of sale of the company within GE Healthcare HiTrap, MBPTrap, StrepTrap, PrimeView, Sephacryl, Super- which supplies them. A copy of these terms and conditions dex, Superloop, ÄKTA and ÄKTAprime are trademarks of...

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