• Exhibits high affinity for his-tagged proteins
• No copurification of proteins
• Resists metal leakage
• Performs well under a wide range of purification conditions
• Cited in over 2,400 peer-reviewed publications

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.

TALON resin can be used in a variety of formats, including FPLC and gravity-flow columns, and small- or large-scale batches. TALON resin is available as a 50% slurry or in pre-packed cartridges and columns. 

If you would like a complete system for bacterial expression and purification, please click here.

Reactive Core Contains Cobalt for Highest Purity

TALON Metal Affinity Resin is complexed with cobalt ions that make it highly selective for polyhistidine-tagged proteins (1–3). 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 reult, TALON resin binds more specifically to polyhistidine-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 ad 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 Superflow™ Resin for FPLC Applications

TALON Superflow is specifically designed for quick and effective purification of polyhistidine-tagged proteins at high flow rates and medium pressure (up to 150 psi)—it’s ideal for FPLC applications. For pre-packed 1 ml and 5 ml cartridges, please visit HisTALON Cartridge.

TALON CellThru Resin for Purification of Membrane-Bound Proteins

TALON CellThru is a novel IMAC resin for purifying polyhistidine-tagged proteins from crude cel lysates, sonicates, and fermentation harvests. This resin uses the same proprietary ligand as our TALON Metal Affinity Resin, but has larger beads (300–500 μm) which permit cellular debris to flow through without centrifugation (Figure 5). TALON CellThru Resin captures polyhistidine-tagged protein directly from crude lysates in one quick step (Figure 6), minimizing protein degradation and generating higher yields of purified protein than conventional strategies.

TALON CellThru Resin is ideal for purifying membrane-bound proteins or multi-protein complexes, which typically pellet with the cell debris during the lysate clarification step and so are not available for binding to the resin. Because crude, non-clarified lysates can be applied directly to the resin, you can skip the centrifugation step, allowing membrane-bound proteins and multi-protein complexes to remain in the lysate where they can bind to the resin. The resin consists of large agarose beads that can be packed into standar chromatography columns with large-pore frits to prevent column blockage. Particulate material flows between the beads while the soluble protein product is captured by the ion exchange functional groups attached to the beads. The resin is ideal for use in expanded bed chromatography for recovering even higher amounts of protein and is recommended for applications such as high-throughput purification of polyhistidine-tagged proteins from crude extracts. We also offer CellThru 10 ml Disposable Columns (with a filter pore size of 90–130 μm) that allow cellular debris to flow through easily.

TALONspin™ Columns

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

TALON Purification Kit

Start here when using TALON resin in your application: This convenient kit contains TALON Resin, columns, and all the buffers necessary to extract, wash, and elute polyhistidine-tagged proteins. Sufficient buffer is provided for the extraction of ten 1 g bacterial pellets.

xTractor Buffer Kit for Optimized Cell Lysis

This buffer system is optimized for extraction of proteins from fresh or frozen bacterial cells. The xTractor Buffer is useful in extracting proteins over a broad range of molecular weights, including high molecular weight proteins that cannot be extracted unless the membrane is fully disrupted. Lysozyme and DNase I are included to help disrupt membranes and to reduce solution viscosity, respectively. The xTractor Buffer is compatible with all TALON resins, allowing quick purification of polyhistiine-tagged proteins.

HisTALON™ Buffer Set

The HisTALON Buffer Set (Cat. No. 635651) is specifically designed for isolation and purification of his-tagged proteins from a variety of samples. It provides a sufficient supply of buffers for 20 purifications (1 ml columns).

The HisTALON Buffer Set provides the following improvements over the TALON Buffer Kit (Cat. No. 635515), which it replaces:

1. Ready to use – Included buffers are all 1X, unlike the previous kit, which contained 5X buffers.
2. Lysis buffer included – The kit comes with xTractor Buffer, which is optimized for TALON resins.
3. Versatile – Improved buffer composition makes the kit versatile enough to be used across all platforms, from gravity flow columns to FPLC cartridges.

Note: TALON 2 ml Disposable Gravity Columns (Cat. No. 63566), do not contain resin.

We highly recommend using Protease Inhibitor Cocktail – ProteoGuard™ with all our protein purification products.

 

 

 

 

Figure 1. The cobalt ion has higher affinity and specificity for polyhistidine-tagged proteins. The indicated metals were immobilized onto sepharose CL-6B (Pharmacia) using TALON Metal Affinity Resin containing a unique tetradentate chelator. 20 μl of eluate from the indicated resin was electrophoresed on a 12% polyacrylamide gel and stained with Coomassie blue.

 

 

Figure 2. Purification of 6xHis-GFPuv under denaturing conditions. The fusion protein was purified in 8M urea using TALON Resin. M: moleculr weight marker.

 

 

Figure 3. Purification, yields and SDS-PAGE fractions of protein purified with TALON Resin in the presence of β-mercaptoethanol. Purification with TALON Superflow Resin requires less washing with exceptional results. Panel A. Native purification of 6xHis protein. 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 β-mercaptoethanol. Even lanes: 20 μl of non-adsorbed material. Odd lanes: 5 μl of eluate. Panel B. Yields of recombinant 6xHis DHFR from cell extracts purified by TALON Superflow Resin versus Nickel-NTA in the presence of βME. N-terminal 6xHis DHFR was expressed and purified under native conditions. Protein concentrations were determined by Bradford assay. Yields are expressed as a percetage of total protein in the cell lysate. Panel C. SDS-PAGE of FPLC fractions from 6xHis-GFPuv purification. FPLC purification fractions were taken from the peaks in Figure 4.

 

 

Figure 4. FPLC purification of 6xHis-GFPuv with TALON Superflow. Nickel-NTA (Panel A) requires longer washing and lower flow rates to purify 6xHis- GFPuv than TALON Superflow Resin (Panel B). Protein was extracted in 50 mM sodium phosphate, 0.3 M NaCl, pH 7.0. Panel A. 3.2 ml culture filtrate was loaded at 0.5 ml/min. Then nonadsorbed material was washed in the same buffer with 10 mM imidazole. Protein was eluted with 20 mM imidazole (Peak II) and 250 mM imidazole (Peak III). Panel B. 3.2 ml culture filtrate was loaded at 1 ml/min. Then, nonadsorbed material was washed with the same extraction buffer and eluted with 150 mM imidazole (Peak I).

 

 

Figure 5. TALON CellThru Resin purifies proteins from crude cell lysates more rapidly than the conventional method.

 

 

Figure 6. 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.

 

 

 

 

 

 

Applications Polyhistidine-tagged protein purification Recombinant protein purification & identification Batch scale protein purification FPLC purification Gravity flow	References 1. Chaga, G. S., et al. (1999) Biotechnol. Appl. Biochem. ,tel:400-650-8506(1):19–24. 2. Porath, J. (1992) Protein Express. Purif. 3(4):263–281. 3. Yang, T., et al. (1997) Amer. Biotechnol. Lab., pp. 12–14. 4. Protein Purification Products Overview (2001) Clontechniques XVI(1):27–32.
Components TALON Purification Kit Components Equilibration Buffer Equilibration/Wash Buffer Elution Buffer TALON Metal Affinity Resin 2-ml Disposable Columns 10-ml Disposable Column TALON xTrator Buffer Kit Components 200 ml xTractor Buffer DNase I Lysozyme	Storage Conditions 4°C for TALON Resin, TALON CellThru Resin, TALON Superflow Resin & TALONspin Columns Room temperature for TALON Disposable Columns & Buffers