GST•Tag Fusion Protein Purification Products  |
• BugBuster® GST•Bind™ Purification Kit
• PopCulture® GST•Mag™ Purification Kit
• RoboPop™ GST•Mag™ Purification Kit
• RoboPop™ GST•Bind™ Purification Kit
 Back to Purification His•Tag and GST•Tag Protein Purification and Detection |
BugBuster GST•Bind™ Purification Kit
Affinity purification of GST fusion proteins |
The BugBuster GST•Bind Purification Kit combines the GST•Bind Resin, GST•Bind Buffer Kit reagents and BugBuster Protein Extraction Reagent for convenient preparation of soluble cell extracts and affinity purification of GST•Tag™ fusion proteins. BugBuster Protein Extraction Reagent is formulated for the gentle disruption of the cell wall of E. coli, resulting in the liberation of soluble protein. Cells are harvested by centrifugation as usual, followed by suspension in BugBuster Reagent. During a brief incubation, soluble proteins are released. The extract is clarified by centrifugation, which removes cell debris and insoluble proteins. The clarified extract is ready to apply to GST•Bind Resin.
Custom Configurations
BugBuster Protein Extraction Reagent and GST•Bind Resin can be custom packaged in larger quantities to suit your needs.
| | Product:
BugBuster GST•Bind Purification Kit
70794-3
Available seperately:
Chromatography Columns
69673-3
Column Extension Funnels
71030-3
BugBuster Protein Extraction Reagent
70584
Benzonase Nuclease, Purity > 90%
70746
PopCulture Reagent
71092
rLysozyme Solution
71110
GST•Bind Resin
70541
GST•Bind Buffer Kits
70534-3
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GST•Bind Purification
A crude extract containing excess unfused GST was applied to a 2-ml GST•Bind Resin column. Total protein yield after purification was 8 mg/ml resin. |
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PopCulture GST•Mag™ Purification Kit
PopCulture extraction and GST•Tag™ fusion protein purification from E. coli cultures |
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 The PopCulture GST•Mag Purification Kit is designed for purification of GST•Tag fusion proteins directly from E. coli cultures without harvesting cells. The procedure combines PopCulture total culture extraction with magnetic affinity purification using GST•Mag Agarose Beads.
PopCulture Reagent is a detergent-based concentrate that can be added directly to cultures of E. coli to effectively extract proteins without the need for centrifugation. Use of GST•Mag Agarose Beads enables the entire procedure to be carried out in a single tube without using columns or centrifugation.
The PopCulture GST•Mag Purification Kit combines PopCulture Reagent, GST•Mag Agarose Beads, corresponding buffers and rLysozyme™ Solution. This kit enables processing of 40 × 3-ml cultures with yields up to 150 μg GST•Tag fusion protein per 3-ml culture, based on bead binding capacity. The kit is compatible with the Novagen Magnetight Separation Stand (see in this chapter). For 96-well processing using PopCulture, please refer to the various RoboPop Purification Kit descriptions onour website.
| | Product:
PopCulture GST•Mag Purification Kit
71113-3 Available seperately:
Benzonase Nuclease, Purity > 90%
70746
Benzonase Nuclease HC, Purity > 90%
71205-3
PopCulture Reagent
71092
rLysozyme Solution
71110
GST•Mag™ Agarose Beads
71084
Magnetight™ HT96™ Stand
71101-3
Magnetight™ Seperation Stand
69964-3
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Features
• No need to separate cells from culture media
• No need to mechanically disrupt cells
• No need to clarify cell extracts prior to purification
• Direct affinity adsorption of target proteins to resin from the total culture extract
• Ability to rapidly perform the entire cell growth and purification process in a single tube or well.
Custom Configurations
PopCulture Reagent, rLysozyme Solution, and GST•Mag Agarose Beads can be custom packaged in larger quantities to suit your needs.
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PopCulture GST•Mag purification
Induced cultures of E. coli strain BL21(DE3) containing pET-41b(+), which encodes a GST•Tag/His•Tag fusion protein, were processed using PopCulture Reagent and GST•Mag Agarose Beads. Samples of a crude extract prepared with BugBuster Reagent and the purified proteins were analyzed by SDS-PAGE and Coomassie blue staining. |
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RoboPop GST•Mag Purification Kit
PopCulture extraction and GST•Mag purification in a 96-well plate |
 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 Sterile 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 the Novagen Magnetight HT96 Separation Stand, which is recommended for efficient processing of magnetic affinity supports in deep-well plates. The Sterile 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 with the Genesis® Freedom™ Workstation from Tecan and the MultiPROBE® II liquid handling work station from PerkinElmer Life Sciences. | | Product:
RoboPop GST •Mag Purification Kit
71102-3 Available seperately:
PopCulture Reagent
71092
Benzonase Nuclease, Purity > 90%
70746
Benzonase Nuclease HC, Purity > 90%
71205-3
rLysozyme Solution
71110
GST•Mag™ Agarose Beads
71084
GST•Bind Buffer Kits
70534-3
Magnetight™ HT96™ Stand
71101-3
Sterile 96-well Deep Well Culture Plate with Sealers
71111-3
Chromatography Columns
69673-3
Column Extension Funnels
71030-3
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Custom Configurations
PopCulture Reagent, rLysozyme Solution, and GST•Mag Agarose Beads can be custom packaged in larger quantities to suit your needs.
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Magnetic-based affinity purification
Cultures of E. coli strain BL21(DE3) containing the vector constructs described in the table were grown at 30°C for 16 h with target protein expression induced by Overnight Express™ Autoinduction System 1. The cultures were processed robotically (Tecan Genesis® 200) according to the RoboPop magnetic-based purification protocol. Lane M: Perfect Protein™ Markers, 10–225 kDa. |
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RoboPop GST•Bind Purification Kit
High-throughput, milligram-scale purification of GST•Tag fusion proteins |
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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 with the Genesis Freedom Workstation from Tecan and the Packard-brand MultiPROBE II liquid handling work station from PerkinElmer Life Sciences. A 96-well Collection Plate (1-ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins.
See the Processing Protocol for the RoboPop GST•Bind Kit
| | Product:
RoboPop GST•Bind Purification Kit
71189-3 Available seperately:
PopCulture Reagent
71092
Benzonase Nuclease, Purity > 90%
70746
Benzonase Nuclease HC, Purity > 90%
71205-3
rLysozyme Solution
71110
GST•Bind Resin
70541
Magnetight™ HT96™ Stand
71101-3
2-ml 96-well Filter Plate
71196-3
Chromatography Columns
69673-3
Column Extension Funnels
71030-3
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Custom Configurations
PopCulture Reagent, rLysozyme Solution, and GST•Bind Resin can be custom packaged in larger quantities to suit your needs. |
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Filtration-based affinity purification
Cultures of E. coli strain BL21(DE3) containing the vector constructs described in the table were grown at 30°C for 16 h with target protein expression induced by Overnight Express™ Autoinduction System 1. The cultures were processed robotically (Tecan Genesis 200) according to the RoboPop filtration-based purification protocol. Lane M: Perfect Protein™ Markers, 10–225 kDa. |
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