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Home › Products › Protein research › Purification products › His-tagged protein purification › Bulk resins and gravity columns › Cobalt columns—spin

His-tagged protein purification

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Fast, small-scale his-tagged protein purification—TALON Spin Columns

TALON Spin Columns contain 0.5 ml of TALON-NX, an immobilized metal affinity chromatography resin for the purification of recombinant his-tagged proteins under native or denaturing conditions.

TALON Spin Columns contain 0.5 ml of TALON-NX, an immobilized metal affinity chromatography resin for the purification of recombinant his-tagged proteins under native or denaturing conditions.

TALON His-Tag Purification Resin lets you prepare exceptionally pure his-tagged proteins from bacterial, mammalian, yeast, and baculovirus-infected cells, under native or denaturing conditions. TALON is an immobilized metal affinity chromatography (IMAC) resin charged with cobalt, which binds to his-tagged proteins with higher specificity than nickel-charged resins. As a result, TALON resin delivers his-tagged proteins of the highest purity. In addition, each cobalt ion is bound to the resin at four sites, resulting in low metal-ion leakage.

Reactive core contains cobalt for highest purity

TALON Metal Affinity Resin is complexed with cobalt ions that make it highly selective for his-tagged proteins. TALON’s cobalt-containing reactive core has strict spatial requirements—only proteins containing adjacent histidines or specially positioned, neighboring histidines are able to bind. The spatial requirements for nickel-based resins (e.g., Ni-NTA) are less strict—these resins have a much higher affinity for randomly positioned (i.e., non-his-tag) histidines. As a result, TALON resin binds more specifically to his-tagged proteins.

Uniform matrix guarantees less contamination

Cobalt-based resins have a more uniform structure than nickel-based resins. TALON resin contains negatively charged reactive sites that form three-dimensional pockets. Each pocket contains three carboxyl groups and one nitrogen atom that collectively bind a single, positively charged cobalt ion—an arrangement that allows the cobalt ion to bind to two adjacent histidine residues. In this configuration, cobalt is bound very tightly and does not leach out of the resin. Nickel-based resins are less homogeneous in structure because nickel ions can form two different coordination complexes: one of which forms a three-dimensional pocket similar to that of the TALON ligand, and a second that forms a planar (flat) structure. In the distorted, planar structure, each nickel ion binds to only two carboxyl groups and one nitrogen atom. As a result, the planar structure binds the nickel ions less tightly, allowing them to leach from the resin. TALON Metal Affinity Resin, with its uniform matrix, provides high affinity binding under a variety of purification conditions, ensuring optimal protein purification.

TALON Spin Columns

These ready-made spin columns contain TALON-NX Resin for the simultaneous purification of several his-tagged proteins in parallel in only 30 minutes. They are recommended for small-scale, single-use applications, such as verifying his-tagged protein expression in transformants, or trial-level purification protocols.

Choice of native or denaturing purification conditions

TALON Resin retains its protein-binding specificity and yield under a variety of purification conditions. It is stable under both denaturing and native (nondenaturing) conditions. Deciding whether to use native or denaturing purification conditions depends on protein location, solubility, accessibility of the his tag, downstream applications, and preservation of biological activity.

  • Native conditions
    Purifying a protein under native conditions is the most efficient way to preserve its biological activity, but requires that the protein be soluble. Advantages include:
    • Eliminating the renaturation step at the end of the purification, saving time, and preventing significant loss of activity
    • Retaining the ability to copurify enzyme subunits, cofactors, and associated proteins
  • Denaturing conditions
    Because proteins that are overexpressed in prokaryotic systems sometimes form insoluble aggregates called inclusion bodies, you may need to purify proteins under denaturing conditions—using strong denaturants such as 6 M guanidinium or 8 M urea to enhance protein solubility. Advantages include:
    • Complete solubilization of inclusion bodies and his-tagged proteins
    • Improved binding to the matrix and reduced nonspecific binding, due to full exposure of the his tag

His-tagged proteins purified under denaturing conditions can be used directly in subsequent applications, or may need to be renatured and refolded. Protein renaturation and refolding can be performed prior to elution from the column. However, yields of recombinant proteins will be lower than under native conditions, because urea and guanidinium molecules compete with histidines for binding to metal.

Use of reducing agents

Purification with TALON Resin may be carried out in the presence of β-mercaptoethanol, but not DTT or DTE, to preserve reduced sulfhydryl (-SH) groups that are important for the biological activity and structure of a given protein. TALON provides higher yields than Ni-NTA in the presence of β-mercaptoethanol.

 More  Less
Cat. # Product Size Price License Quantity Details
635603 TALON® Spin Columns 50 x 0.5 mL Columns $428.00

License Statement

ID Number  
76 The use of TALON Purification products is covered under U.S. Patent No. 5,962,641. For-Profit and Not-For-Profit purchasers of TALON Purification products are entitled to use the reagents for internal research only. For information on licensing TALON Chemistry for commercial purposes, including but not limited to use in manufacturing, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

Spin columns each containing 0.5 ml of TALON-NX, an immobilized metal affinity chromatography resin for the purification of recombinant his-tagged proteins under native or denaturing conditions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

SDS-PAGE of TALON CellThru Resin purified proteins

SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.

Back

Native vs. denaturing purification procedures using TALON resin

Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.

Back

Native purification with TALON resin preserves biological activity of proteins

Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.

Back

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.

Back

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol
Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol. N-terminal 6xHis-tagged mouse DHFR (19.5 kDa) was expressed in E. coli. 2 ml of lysate was purified using gravity flow on TALON resin in increasing concentrations of beta-mercaptoethanol. Even lanes: 20 μl of nonadsorbed material. Odd lanes: 5 μl of eluate

Back

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin
Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin. N-terminal 6xHis DHFR was expressed and purified under native conditions. Protein concentrations were determined by Bradford assay. Yields are expressed as a percentage of total protein in the cell lysate.

Back

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification
TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification. Rapid, parallel purification in as little as 30 minutes.
635602 TALON® Spin Columns 25 x 0.5 mL Columns $264.00

License Statement

ID Number  
76 The use of TALON Purification products is covered under U.S. Patent No. 5,962,641. For-Profit and Not-For-Profit purchasers of TALON Purification products are entitled to use the reagents for internal research only. For information on licensing TALON Chemistry for commercial purposes, including but not limited to use in manufacturing, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

Spin columns each containing 0.5 ml of TALON-NX, an immobilized metal affinity chromatography resin for the purification of recombinant his-tagged proteins under native or denaturing conditions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

SDS-PAGE of TALON CellThru Resin purified proteins

SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.

Back

Native vs. denaturing purification procedures using TALON resin

Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.

Back

Native purification with TALON resin preserves biological activity of proteins

Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.

Back

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.

Back

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol
Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol. N-terminal 6xHis-tagged mouse DHFR (19.5 kDa) was expressed in E. coli. 2 ml of lysate was purified using gravity flow on TALON resin in increasing concentrations of beta-mercaptoethanol. Even lanes: 20 μl of nonadsorbed material. Odd lanes: 5 μl of eluate

Back

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin
Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin. N-terminal 6xHis DHFR was expressed and purified under native conditions. Protein concentrations were determined by Bradford assay. Yields are expressed as a percentage of total protein in the cell lysate.

Back

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification
TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification. Rapid, parallel purification in as little as 30 minutes.
635601 TALON® Spin Columns 10 x 0.5 mL Columns $141.00

License Statement

ID Number  
76 The use of TALON Purification products is covered under U.S. Patent No. 5,962,641. For-Profit and Not-For-Profit purchasers of TALON Purification products are entitled to use the reagents for internal research only. For information on licensing TALON Chemistry for commercial purposes, including but not limited to use in manufacturing, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

Spin columns each containing 0.5 ml of TALON-NX, an immobilized metal affinity chromatography resin for the purification of recombinant his-tagged proteins under native or denaturing conditions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

SDS-PAGE of TALON CellThru Resin purified proteins

SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.

Back

Native vs. denaturing purification procedures using TALON resin

Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.

Back

Native purification with TALON resin preserves biological activity of proteins

Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.

Back

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin

Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.

Back

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol

Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol
Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol. N-terminal 6xHis-tagged mouse DHFR (19.5 kDa) was expressed in E. coli. 2 ml of lysate was purified using gravity flow on TALON resin in increasing concentrations of beta-mercaptoethanol. Even lanes: 20 μl of nonadsorbed material. Odd lanes: 5 μl of eluate

Back

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin

Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin
Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin. N-terminal 6xHis DHFR was expressed and purified under native conditions. Protein concentrations were determined by Bradford assay. Yields are expressed as a percentage of total protein in the cell lysate.

Back

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification

TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification
TALON Spin column: Ready-to-use prepacked spin columns for small-scale his-tag purification. Rapid, parallel purification in as little as 30 minutes.

Overview

  • Exhibits high affinity for his-tagged proteins
  • No copurification of proteins
  • Resists metal leakage
  • Performs well under a wide range of purification conditions
  • Ready-to-use spin columns
  • Each column contains 0.5 ml of resin

More Information

Applications

  • His-tagged protein purification
  • Recombinant protein purification & identification
  • Spin column purification

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


Featured kits Reagent compatibility
Overviews Native vs denaturing purifications
Technical notes See the data: TALON resins

TALON resin application protocols

Denaturing purification

Choose between mini-scale purification, batch/gravity-flow column purification, and large-scale purification protocols.


Why choose TALON resin?

Obtain highest purity with cobalt resin

Prepare exceptionally pure his-tagged proteins from bacterial, mammalian, yeast, and baculovirus-infected cells.

See the data

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Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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