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HisPur Ni-NTA Resin and Kits


High-capacity nickel-IMAC resin for His-tagged fusion protein purification.

HisPur Ni-NTA Resin for high-yield 6xHis protein purification

Thermo Scientific HisPur Ni-NTA Resin is a high-capacity, high-performance nickel-IMAC resin for routine affinity purification of His-tagged fusion proteins.

The specially prepared support consists of beaded agarose derivatized with the nitrilotriacetic acid (NTA) chelation moiety and loaded with divalent nickel ions (Ni2+). The immobilized metal affinity chromatography (IMAC) resin provides exceptional binding capacity and performance for recombinant His-tagged protein purification.

HisPur Ni-NTA Resin is available in several package sizes of resin slurry, three sizes of centrifuge-ready columns, complete purification kits, two sizes of FPLC-ready chromatography cartridges, and 96-well filter plates for high through-put needs. With these options, high-yield His-tag protein purification is possible at nearly any scale.

Highlights:

  • High capacity – bind up to 60mg of 6xHis-tagged protein per milliliter of resin
  • Versatile – purify proteins using native or denaturing conditions
  • Compatible – use with Thermo Scientific Cell Lysis Reagents and a variety of buffer additives
  • Flexible – available in multiple formats including bulk resin, spin columns, chromatography cartridges and 96-well filter plates
  • Cost-effective – reuse the same batch of resin at least five times
  • Easy to use – pre-formulated buffers available for kit formats

Available Formats:

Product Details:

The expression and purification of recombinant proteins is central to protein regulation, structure and function studies. The majority of recombinant proteins are expressed as fusions with short affinity tags, the most popular being the polyhistidine (6xHis) tag. The method used to purify recombinant His-tagged proteins is immobilized metal affinity chromatography (IMAC), consisting of chelating resins charged with either nickel or cobalt ions that coordinate with the histidine side chains.

HisPur Ni-NTA Resin effectively purifies high levels of overexpressed His-tagged fusion proteins from bacterial lysates, such as those that are attained with Thermo Scientific B-PER Bacterial Protein Extraction Reagents. The resin performs well in batch-binding and spin-column procedures at a variety of scales. Performance equals or exceeds popular Ni-NTA resins from other suppliers.

HisPur Ni-NTA Resin is effective in batch and spin column protocols
Thermo Scientific HisPur Ni-NTA resin is effective for purifying His-tagged proteins in batch-bind and spin-column formats with comparable results to another supplier's resin. Left panel: Bacterial lysate (10mg total protein) containing over-expressed 6xHis-annexin V was applied to 0.2mL of HisPur Ni-NTA Resin and purified by the batch-bind method. Center panel: Bacterial lysate (1.7mg total protein) containing over-expressed 6xHis-annexin V was applied to a HisPur Ni-NTA Spin Column (0.2mL) and purified by the spin method. Right panel: The same amount of lysate (1.7mg) was applied to Supplier Q's spin column and purified per the manufacturer's instructions. Gels lanes were normalized to equivalent volume. M = molecular-weight marker, L = lysate load, FT = flow-through and E = elution.

 

HisPur Ni-NTA resin is better than other nickel IMAC products

Thermo Scientific HisPur Ni-NTA resin performs as well or better than other suppliers' nickel resins. Bacterial lysate (12mg total protein) containing over-expressed 6xHis-green fluorescent protein (GFP) was applied to HisPur Ni-NTA Resin (0.2mL) and purified by the batch-bind method. The same amount of total protein was applied to Supplier Q, Supplier C, and Ni-IDA resins per the manufacturers' instructions. Gel lanes were normalized to equivalent volume. M = molecular-weight marker and L = lysate load.

HisPur Ni-NTA Resin is a high-quality, stable and resilient affinity support. Tests confirm that no decrease in performance occurs after at least five repeated uses. These data indicated that the resin is highly resistant to structural degradation or nickel ion leaching during normal use.

HisPur Ni-NTA Resin is reusable
Thermo Scientific HisPur Ni-NTA Resin can be used at least five times without losing performance. Bacterial lysate (20mg total protein) containing over-expressed 6xHis-Protein L was applied to HisPur Ni-NTA Resin (0.2mL) and purified by the batch-bind method. Before each reuse, the resin was washed with 20mM MES buffer, 0.1M NaCl; pH 5 (1mL) and followed by a water wash (1mL). Gels lanes were normalized to equivalent volume. M = MW marker, L = lysate load, FT = flow-through and E = elution.

HisPur Ni-NTA Resin and HisPur Cobalt Resin are alternative forms of IMAC, the former using nickel and the latter using cobalt as the chelated metal ion responsible for His-tag binding. Ni-NTA resins have been the most common IMAC resin choice for 6xHis-tag protein purifications because of the four metal-binding sites on the chelate, which enables high-protein binding and low-metal ion leaching. Cobalt binds less strongly and therefore more discriminantly, enabling greater purity but usually less yield in His-tagged protein purification.

HisPur Nickel and Cobalt Resins maximize yield and purity respectively
HisPur Ni-NTA and HisPur Cobalt Resins maximize yield and purity, respectively. Gel panels: Bacterial lysate containing over-expressed 6xHis-AIF2 (6mg total protein) or 6xHis-GFP (4mg total protein) was applied to HisPur Ni-NTA Resin (0.2mL) and purified by the batch-bind method. The same amount of total protein was applied to Ni-IDA and HisPur Cobalt Resins and purified as described in the instructions. Gels lanes were normalized to equivalent volume. M = molecular-weight marker, L = lysate load and FT = flow-through.
Table. Elution fractions were analyzed for protein content using the Coomassie Plus (Bradford) Protein Assay Kit (Part No. 23236). Purity was determined by analyzing the gel with densitometry software.
  6xHis AIF2 (73kDa) 6xHis GFP (28kDa)
Resin Yield Purity Yield Purity
HisPur Ni-NTA 0.5mg 32% 0.8mg 90%
Ni-IDA 0.5mg 25% 0.6mg 52%
HisPur Cobalt 0.4mg 49% 0.7mg 91%

Related Resources:

Review of His-tagged fusion protein production and purification

Related Products:

His-Tag Antibodies
His-tagged protein purification products – including Ni-NTA and Cobalt Superflow resins
Ni-NTA Magnetic Beads – benchtop or automated magnetic separations
Nickel Coated Plates – for plate-based assays with His-tagged proteins
B-PER Bacterial Protein Extraction Reagents – for high-yield extraction from bacteria
Glutathione and HA-tag purification resins – for fusion protein purification
Fisher BioReagents Imidazole


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
88221 HisPur Ni-NTA Resin
Formulation: Nickel-NTA (IMAC) on crosslinked beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding up to 60mg of His-tagged protein per mL of resin
10mL Product Instructions for product #88221 HisPur Ni-NTA Resin MSDS for product #88221 HisPur Ni-NTA Resin $89.00

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88222 HisPur Ni-NTA Resin
Formulation: Nickel-NTA (IMAC) on crosslinked beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding up to 60mg of His-tagged protein per mL of resin
100mL Product Instructions for product #88222 HisPur Ni-NTA Resin MSDS for product #88222 HisPur Ni-NTA Resin $605.00

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88223 HisPur Ni-NTA Resin
Formulation: Nickel-NTA (IMAC) on crosslinked beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding up to 60mg of His-tagged protein per mL of resin
500mL Product Instructions for product #88223 HisPur Ni-NTA Resin MSDS for product #88223 HisPur Ni-NTA Resin $2,450.00

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88224 HisPur Ni-NTA Spin Columns, 0.2mL
Formulation: 0.8mL-capacity microcentrifuge columns containing 0.2mL resin
Sufficient For: Binding up to 12mg of His-tagged protein per column
25 columns Product Instructions for product #88224 HisPur Ni-NTA Spin Columns, 0.2mL MSDS for product #88224 HisPur Ni-NTA Spin Columns, 0.2mL $163.00

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88225 HisPur Ni-NTA Spin Columns, 1mL
Formulation: 8mL-capacity centrifuge columns containing 1mL resin
Sufficient For: Binding up to 60mg of His-tagged protein per column
5 columns Product Instructions for product #88225 HisPur Ni-NTA Spin Columns, 1mL MSDS for product #88225 HisPur Ni-NTA Spin Columns, 1mL $104.00

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88226 HisPur Ni-NTA Spin Columns, 3mL
Formulation: 22mL-capacity centrifuge columns containing 3mL resin
Sufficient For: Binding up to 180mg of His-tagged protein per column
5 columns Product Instructions for product #88226 HisPur Ni-NTA Spin Columns, 3mL MSDS for product #88226 HisPur Ni-NTA Spin Columns, 3mL $162.00

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88227 HisPur Ni-NTA Spin Purification Kit, 0.2mL
Formulation: Multi-component kit
Sufficient For: Purifying up to 12mg of His-tagged protein per column

Kit Contents:
HisPur Ni-NTA Spin Columns, 0.2mL, 25 columns
Phosphate-buffered Saline (10X), 30mL
Imidazole (2M), 8mL
Collection Tubes, 80 tubes
Column Plugs, 25 plugs
25-column kit Product Instructions for product #88227 HisPur Ni-NTA Spin Purification Kit, 0.2mL MSDS for product #88227 HisPur Ni-NTA Spin Purification Kit, 0.2mL $282.00

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88228 HisPur Ni-NTA Spin Purification Kit, 1mL
Formulation: Multi-component kit
Sufficient For: Purifying up to 60mg of His-tagged protein per column

Kit Contents:
HisPur Ni-NTA Spin Columns, 1mL, 5 columns
Phosphate-buffered Saline (10X), 30mL
Imidazole (2M), 8mL
Column Plugs, 5 plugs
5-column kit Product Instructions for product #88228 HisPur Ni-NTA Spin Purification Kit, 1mL MSDS for product #88228 HisPur Ni-NTA Spin Purification Kit, 1mL Certificate of Analysis for product #88228 HisPur Ni-NTA Spin Purification Kit, 1mL $265.00

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88229 HisPur Ni-NTA Spin Purification Kit, 3mL
Formulation: Multi-component kit
Sufficient For: Purifying up to 180mg of His-tagged protein per column

Kit Contents:
HisPur Ni-NTA Spin Columns, 3mL, 5 columns
Phosphate-buffered Saline (10X), 30mL
Imidazole (2M), 25mL
Column Plugs, 5 plugs
5-column kit Product Instructions for product #88229 HisPur Ni-NTA Spin Purification Kit, 3mL MSDS for product #88229 HisPur Ni-NTA Spin Purification Kit, 3mL $282.00

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90098 HisPur Ni-NTA Chromatography Cartridges, 1mL
Formulation: 1mL resin cartridges with LC and Luer-Lok Fittings
Sufficient For: Binding up to 60mg of His-tagged protein per cartridge

Learn more about Pierce Chromatography Cartridges.
5 cartridges Product Instructions for product #90098 HisPur Ni-NTA Chromatography Cartridges, 1mL MSDS for product #90098 HisPur Ni-NTA Chromatography Cartridges, 1mL Certificate of Analysis for product #90098 HisPur Ni-NTA Chromatography Cartridges, 1mL $108.00

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90099 HisPur Ni-NTA Chromatography Cartridges, 5mL
Formulation: 5mL resin cartridges with LC and Luer-Lok Fittings
Sufficient For: Binding up to 300mg of His-tagged protein per cartridge

Learn more about Pierce Chromatography Cartridges.
2 cartridges Product Instructions for product #90099 HisPur Ni-NTA Chromatography Cartridges, 5mL MSDS for product #90099 HisPur Ni-NTA Chromatography Cartridges, 5mL Certificate of Analysis for product #90099 HisPur Ni-NTA Chromatography Cartridges, 5mL $185.00

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88230 HisPur Ni-NTA Spin Plates
Formulation: 96-well filter plates containing 50┬ÁL of resin per well; Includes 6 collection plates
Sufficient For: Binding approx. 1mg of His-tagged protein per well

Learn more about Pierce 96-well Filter Spin Plates.
2 plates Product Instructions for product #88230 HisPur Ni-NTA Spin Plates MSDS for product #88230 HisPur Ni-NTA Spin Plates Certificate of Analysis for product #88230 HisPur Ni-NTA Spin Plates $323.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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