His•Tag Fusion Protein Purification Products  |
• BugBuster® Ni-NTA His•Bind® and His•Bind Purification Kits
• PopCulture® His•Mag™ Purification Kit Purification Kits
• RoboPop™ His•Mag™ Purification Kit
• RoboPop™ Ni-NTA His•Bind® Purification Kit
• Insect RoboPop™ Ni-NTA His•Bind® Purification Kit
 Back to Purification His•Tag and GST•Tag Protein Purification and Detection |
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BugBuster Ni-NTA His•Bind and His•Bind Purification Kits
Convenient preparation of extracts and affinity purification of His•Tag® fusion proteins
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The BugBuster Ni-NTA His•Bind and BugBuster His•Bind Purification Kits combine Ni-NTA His•Bind or His•Bind Resin, respectively, His•Bind Buffer Kit (His•Bind Kit only), Benzonase Nuclease, and BugBuster Protein Extraction Reagent for convenient preparation of soluble cell extracts and affinity purification of His•Tag fusion proteins. BugBuster Protein Extraction Reagent, when combined with rLysozyme™ Solution, gently disrupts the cell wall of E. coli to liberate cellular proteins.
In practice, cells are harvested by centrifugation and suspended in BugBuster Reagent. At this point, Benzonase Nuclease can be added to reduce the viscosity of the extract due to liberation of chromosomal DNA. After a brief incubation, insoluble cell debris and inclusion bodies are removed by centrifugation. The clarified extract is ready to use and fully compatible with Ni-NTA His•Bind and His•Bind Resins. Following binding to affinity resin, excess BugBuster is easily removed by washing the column with the appropriate buffer.
Use BugBuster Ni-NTA His•Bind Purification Kit for the purification of proteins in a reducing environment (Ni-NTA His•Bind Resin is compatible with up to 20 mM b-Mercaptoethanol). Use BugBuster His•Bind Purification Kit if you are planning to reuse His•Bind Resin many times.
Custom Configurations
BugBuster Protein Extraction Reagent and His•Bind Resin can be custom packaged in larger quantities to suit your needs. 
| Product:
BugBuster Ni-NTA His•Bind Purification Kit
70751-3
BugBuster His•Bind Purification Kit
70793-3
His•Bind® Purification Kit
70293-3 Available seperately:
BugBuster Protein Extraction Reagent
70584
Benzonase Nuclease, Purity > 90%
70746
Ni-NTA His•Bind®Resin
70666
His•Bind Resin
69670
Ni-NTA Buffer Kit
70899-3
His•Bind Buffer Kit
69755
8X Binding Buffer
69754
8X Wash Buffer
69756
4X Elute Buffer
69757
4X Strip Buffer
69758
8X Charge Buffer
69759
Chromatography Columns
69673-3
Column Extension Funnels
71030-3 |
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A target protein comprising less than 5% of the total protein prepared from a recombinant in pET-32b(+) was purified using His•Bind Resin and His•Bind Buffer Kit. The high specificity of the His•Bind Resin is demonstrated by analysis of the indicated fractions by SDS-PAGE and Coomassie blue staining. |
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PopCulture His•Mag Purification Kit
PopCulture extraction and His•Mag purification from E. coli cultures |
 The PopCulture His•Mag Purification Kit is designed for purification of His•Tag fusion proteins directly from E. coli cultures without harvesting cells. The procedure combines PopCulture total culture extraction with magnetic affinity purification using His•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 His•Mag Agarose Beads enables the entire procedure to be carried out in a single tube without using columns or centrifugation.
The PopCulture His•Mag Purification Kit combines PopCulture Reagent, His•Mag Agarose Beads, corresponding buffers and rLysozyme Solution. This kit enables the processing of 40 × 3-ml cultures with yields up to 375 μg His•Tag fusion protein per 3-ml culture, based on bead binding capacity. The kit is compatible with the Novagen Magnetight Separation Stand. For 96-well processing using PopCulture, please refer to the next page for more information about the RoboPop His•Mag Purification Kits.
| Product:
PopCulture His•Mag Purification Kit
71114-3 Available seperately:
PopCulture Reagent
71092
His•Mag Agarose Beads
71002
8X Binding Buffer
69754
8X Wash Buffer
69756
4X Elute Buffer
69757
rLysozyme Solution
71110
Magnetight HT96 Stand
71101-3
Magnetight Separation Stand
69964-3
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Features
• No need to separate cells from culture medium
• 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 His•Mag Agarose Beads can be custom packaged in larger quantities to suit your needs.
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PopCulture His•Mag Purification
Induced cultures of E. coli strain BL21(DE3) containing pET-41b(+), which encodes a His•Tag fusion protein, were processed using PopCulture Reagent and His•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 His•Mag Purification Kit
PopCulture extraction and His•Mag purification in a 96-well format |
 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 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 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.
| Product:
RoboPop His•Mag Purification Kit
71103-3 Available seperately:
Magnetight HT96 Stand
71101-3
Benzonase Nuclease, Purity > 90%
70746
Benzonase Nuclease HC, Purity > 90%
71205-3
PopCulture Reagent
71092
rLysozyme Solution
71110
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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 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 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. |
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Automated purification of His•Tag fusion proteins
Cultures of E. coli strain BL21(DE3) containing the vector constructs described in the table above 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 purification protocols. Panel A (approximately 1 μg protein load) and panel B (approximately 2 μg protein load) show purified protein samples analyzed by SDS-PAGE (10–20% gradient gel with Coomassie blue staining). Protein assays were performed by the Bradford method and purity was determined by densitometry of the scanned gel. Lane M: Perfect Protein Markers, 10–225 kDa. |
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RoboPop Ni-NTA His•Bind Purification Kit
High-throughput, milligram-scale purification of His•Tag fusion proteins |
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The RoboPop Ni-NTA His•Bind Purification Kit is designed for filtrationbased 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 centrifugefree 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. Whereas 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 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. A 96-well Collection Plate (1-ml wells) with an air-tight aluminum foil sealer is provided for storage of the purified proteins.
| Product:
RoboPop Ni-NTA His•Bind Purification Kit
71188-3 Available seperately:
PopCulture Reagent
71092
Ni-NTA Buffer Kit
70899-3
rLysozyme Solution
71110
Benzonase Nuclease, Purity > 90%
70746
Benzonase Nuclease HC, Purity > 90%
71205-3
2-ml 96-well Filter Plate
71196-3
Chromatography Columns
69673-3
Column Extension Funnels
71030-3
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Features
• High-throughput, 96-well protein purification directly from E. coli without harvesting cells
• Centrifuge-free cell lysis and extract preparation in one step
• Large-scale culture processing—up to 5 ml per well
• High protein yield—up to 1 mg per well
• Validated for robotic sample processing with the Genesis® Freedom™ Workstation from Tecan and the MultiPROBE® II liquid handling work station from PerkinElmer Life Sciences See the Processing Protocol for the RoboPop Ni-NTA His•Bind Purification Kit
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Custom Configurations
PopCulture Reagent, rLysozyme Solution, and His•Mag Agarose Beads 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 purification protocols. Lane M: Perfect Protein Markers, 10–225 kDa. |
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Insect RoboPop Ni-NTA His•Bind® Purification Kit
Filtration-based, 96-well format purification directly from transfected cultures of insect cells |
The Insect RoboPop Ni-NTA His•Bind Purification Kit is designed for filtration based, 96-well format purification of His•Tag fusion proteins directly from transfected insect cell cultures. This kit includes Insect PopCulture Reagent for protein extraction from total cultures, Benzonase Nuclease for viscosity reduction, Ni-NTA His•Bind Resin and buffers, 2-ml 96-well Filter Plate and Collection Plate with Sealer. The kit is configured for robotic processing of transfected 10-ml suspension cultures and purifies up to 400 μg His•Tag fusion protein per culture based on binding capacity of the resin. Protein yields ranging from 60 to 140 μg per 10 ml culture were obtained with b-gal, Fluc, MAP kinase, and cdc2 kinase expressed in the pIEx™ transient protein expression system (1).
In practice, Insect PopCulture Reagent is added directly to the cell culture, followed by the addition of Benzonase Nuclease. The lysate is transferred to a tube of appropriate size or deep-well plate and Ni-NTA His•Bind resin is added. After mixing, the lysate-resin slurry is transferred to the Filter Plate, and washing and elution steps are carried out on a vacuum manifold. The Filter Plate is compatible with standard vacuum manifolds for either manual or automated processing, and the entire purification has been validated for robotic sample processing with the MultiPROBE® II liquid handling workstation from PerkinElmer Life Sciences and with the Genesis® Freedom™ workstation from Tecan.
1. Loomis, K., Grabski, A., Wong, S. C. (2002)
InNovations 15 16–17.
| Product:
Insect RoboPop Ni-NTA His•Bind Purification Kit
71257-3
Available seperately:
BCA Protein Assay Kit
71285-3
Insect PopCulture Reagent
71187
Benzonase Nuclease, Purity > 99%
70664-3
Benzonase Nuclease, Purity > 90%
70746
Ni-NTA His•Bind Resin
70666
Ni-NTA Buffer Kit
70899-3
2-ml 96-well Filter Plate
71196-3
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Also use the Insect RoboPop Ni-NTA His•Bind Purification Kit with centrifugation when a vacuum manifold is not available. 
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| Sf9 cells in 10-ml suspension cultures (1 × 106 cells/ml) were transfected with 20 μg of the indicated pIEx recombinants using Insect GeneJuice Transfection Reagent. Total culture extracts were prepared 48 h later by the addition of Insect PopCulture Reagent (500 μl) followed by the addition of Benzonase Nuclease (5 μl). Samples were taken at this point to represent the total cell protein. Ni-NTA His•Bind Resin (50 μl per culture) was then added to the extracts. Samples were processed robotically using a Genesis Freedom workstation from Tecan. Target protein was eluted in a volume of 150 μl. Ten μl crude and purified fractions were loaded in adjacent lanes of a 10–20% SDS polyacrylamide gel, which was stained with Coomassie blue. Purified protein yields were determined by BCA assay. |
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