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 His•Tag® and GST•Tag™ Fusion Protein Purification in a 96-well Format |
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Novagen offers a variety of high-throughput affinity purification platforms, providing options for rapid magnetic-based separations with protein yield of up to 125 µg per well, or filtration-based methods for purifying up to 1 mg of recombinant protein per well.
| Product | Culture scale | Processing method | Capacity* |
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| RoboPop™ His•Mag® Purification Kit | 96 × 1 ml | Magnetic | 125 µg/culture | | RoboPop™ GST•Mag™ Purification Kit | 96 × 1 ml | Magnetic | 50 µg/culture | | RoboPop™ Ni-NTA His•Bind® Purification Kit | 96 × 5 ml | Filtration | 1 mg/culture | | RoboPop™ GST•Bind™ Purification Kit | 96 × 5 ml | Filtration | 0.8 mg/culture | | * Capacities are based on 1- or 5-ml cultures and binding capacities of the resin. Yields will vary with the expression levels, folding properties, and solubility of individual fusion proteins. |
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| Magnetic-Based Purification Kits |
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| RoboPop His•Mag Purification Kit | RoboPop GST•Mag Purification Kit |
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The RoboPop His•Mag Purification Kit is designed for 96-well format purification of His•Tag fusion proteins directly from E. coli cultures without harvesting cells. The kits feature PopCulture Reagent for extraction of of proteins from total cultures without the need for centrifugation, and His•Mag Agarose Beads for high-capacity magnetic affinity purification. The combination of PopCulture and magnetic agarose beads enables the entire procedure to be carried out in a single culture plate.
The kits contain on 96-well deep well Culture Plate (2 ml wells) with three air-permeable sealing membranes for bacterial cell growth and protein purification, and one 96-well Collection Plate (450 µl wells) with an air-tight aluminum foil sealer for storage of the purified proteins. rLysozyme Solution, Benzonase Nuclease and purification buffers are also included. The Culture Plate is compatible with Novagen's Magnetight™ HT96™ Separation Stand, which is recommended for efficient processing of magnetic affinity supports in deep well plates.
The RoboPop His•Mag Purification Kit will purify up to 12 mg of His•Tag fusion proteins per plate (up to 125 µg/well). Stated yields are based on 1 ml cultures and binding capacities of the beads, and will vary with the folding properties, expression levels, and solubility of individual fusion proteins. The RoboPop His•Mag Purification Kit has been validated for robotic sample processing. | The RoboPop GST•Mag Purification Kit is designed for 96-well format purification of GST•Tag fusion proteins directly from E. coli cultures without harvesting cells. The kit features PopCulture Reagent for extraction of proteins from total cultures without the need for centrifugation, and GST•Mag Agarose Beads for high-capacity magnetic affinity purification. The combination of PopCulture and magnetic agarose beads enables the entire procedure to be carried out in a single culture plate. The kit contains one 96-well Deep Well Culture Plate (2 ml wells) with three air-permeable sealing membranes for bacterial cell growth and protein purification, and one 96-well Collection Plate (450 µl wells) with an air-tight aluminum foil sealer for storage of the purified proteins. rLysozyme Solution, Benzonase Nuclease and purification buffers are also included. The Culture Plate is compatible with NovagenÕs Magnetight HT96 Separation Stand (see page 28), which is recommended for efficient processing of magnetic affinity supports in deep well plates. The 96-well Deep Well Culture Plate with Sealers is available separately.
The RoboPop GST•Mag Purification Kit will purify up to 4.8 mg of GST•Tag fusion proteins per plate (up to 50 µg/well). Stated yields are based on 1 ml cultures and binding capacities of the beads, and will vary with the folding properties, expression levels, and solubility of individual fusion proteins. The RoboPop His•Mag Purification Kit has been validated for robotic sample processing. | | RoboPop™ His•Mag™ Purification Kit | Cat. No. 71103-3
Components: | | • | 15 ml | PopCulture™ Reagent | | • | 1 | Sterile 96-well Deep Well Culture Plate
with Sealers (3) | | • | 1 | Collection Plate with Sealer | | • | 3 × 1 ml | His•Mag Agarose Beads | | • | 80 ml | 8X Binding Buffer | | • | 2 × 25 ml | 8X Wash Buffer | | • | 50 ml | 4X Elute Buffer | | • | 300 KU | rLysozyme™ Solution | | • | 1 ml | rLysozyme Dilution Buffer | | • | 2500 U | Benzonase® Nuclease,
Purity > 90% |
| | RoboPop GST•Mag™ Purification Kit | Cat. No. 71102-3
Components: | | • | 15 ml | PopCulture™ Reagent | | • | 1 | Sterile 96-well Deep Well Culture Plate
with Sealers (3) | | • | 1 | Collection Plate with Sealer | | • | 3 × 1 ml | GST•Mag Agarose Beads | | • | 2 × 100 ml | 10X GST Bind/Wash Buffer | | • | 40 ml | 10X GLutathione Reconstitution Buffer | | • | 1 g | Reduced Glutathione | | • | 300 KU | rLysozyme™ Solution | | • | 1 ml | rLysozyme Dilution Buffer | | • | 2500 U | Benzonase® Nuclease,
Purity > 90% |
| | Filtration-Based Purification Kits |
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| RoboPop Ni-NTA His•Bind Purification Kit | RoboPop GST•Bind Purification Kit |
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The RoboPop Ni-NTA His•Bind Purification Kit is designed for filtration-based 96-well format purification of soluble His•Tag fusion proteins directly from E. coli cultures without harvesting cells. The kit features PopCulture Reagent, rLysozyme™ Solution, and Benzonase® Nuclease for centrifuge-free cell lysis and extract preparation in one step. The combination of PopCulture extraction, Ni-NTA His•Bind Resin, and a 2 ml filter plate allows high-throughput processing of up to 5 ml of E. coli culture per well. Wheras the magnetic-based His•Mag kit purifies up to 125 µg target protein per 1 ml culture, the filtration-based kit purifies up to 1 mg His•Tag fusion protein per 5 ml culture.
Bacterial culture, cell lysis, and resin binding steps are carried out in standard 24-well plates (not supplied), which accommodate a maximum volume of 5 ml per well. The reaction slurry is then transferred to a 96-well Filter Plate (included) and the washing and elution steps are carried out on a vacuum manifold. The Filter Plate is compatible with standard filter manifolds for manual sample processing, and the entire purification has veen validated for robotic sample processing. A 96-well Collection Plate (1 ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins. | The RoboPop GST•Bind Purification Kit is designed for filtration-based 96-well format purification of soluble GST•Tag fusion proteins directly from E. coli cultures without harvesting cells. The kit features PopCulture Reagent, rLysozyme Solution, and Benzonase Nuclease for centrifuge-free cell lysis and extract preparation in one step. The combination of PopCulture extraction, GST•Bind Resin, and a 2 ml filter plate allows high-throughput processing of up to 5 ml of E. coli culture per well. Whereas the magnetic-based GST•Mag kit purifies up to 50 µg target protein per 1 ml culture, the filtration-based kit purifies up to 0.8 mg GST•Tag fusion protein per 5 ml culture.
Bacterial culture, cell lysis, and resin binding steps are carried out in standard 24-well plates (not supplied), which accommodate a maximum volume of 5 ml per well. The reaction slurry is then transferred to a 96-well Filter Plate (included) and the washing and elution steps are carried out on a vacuum manifold. The Filter Plate is compatible with standard filter manifolds for manual sample processing, and the entire purification has been validated for robotic sample processing. A 96-well Collection Plate (1 ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins. | | RoboPop™ Ni-NTA His•Bind® Purification Kit | Cat. No. 71188-3
Components: | | • | 75 ml | PopCulture™ Reagent | | • | 25 ml | Ni-NTA His•Bind Resin | | • | 125 ml | 4X Ni-NTA Bind Buffer | | • | 2 × 125 ml | 4X Ni-NTA Wash Buffer | | • | 50 ml | 4X Ni-NTA Elute Buffer | | • | 1 | 2 ml 96-well Filter Plate | | • | 1 | 1 ml 96-well Collection Plate with Sealer | | • | 300 KU | rLysozyme™ Solution | | • | 1 ml | rLysozyme Dilution Buffer | | • | 10 KU | Benzonase® Nuclease,
Purity > 90% |
| | RoboPop™ GST•Bind™ Purification Kit | Cat. No. 71189-3
Components: | | • | 75 ml | PopCulture™ Reagent | | • | 25 ml | GST•Bind Resin | | • | 100 ml | 10X GST Bind/Wash Buffer | | • | 1 g | Reduced Glutathione | | • | 40 ml | 10X GLutathione Reconstitution Buffer | | • | 1 | 2 ml 96-well Filter Plate | | • | 1 | 1 ml 96-well Collection Plate with Sealer | | • | 300 KU | rLysozyme™ Solution | | • | 1 ml | rLysozyme Dilution Buffer | | • | 10 KU | Benzonase® Nuclease,
Purity > 90% |
| All RoboPop Purification Kits are validated on the Genesis® Freedom™ Workstation from Tecan and on the MultiPROBE® II HT EX Robot from PerkinElmer Life Sciences.
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