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HisPur Cobalt Resin, Columns and Purification Kits


Specific, fast and gentle purification of His-tagged proteins.

HisPur Cobalt Resin for IMAC metal chelate purification of His-tagged proteins

Thermo Scientific HisPur Cobalt Resin is a tetradentate chelating agarose resin charged with divalent cobalt (Co2+) for obtaining high-purity his-tagged proteins with no metal contamination.

HisPur Cobalt Agarose 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-quality His-tag protein purification is possible at nearly any scale. Compared to Ni-IDA and tetradentate Ni2+ chelate resins, HisPur Cobalt Resin binds histidine-tagged proteins with higher specificity (less off-target binding) and releases them with gentler conditions (lower concentrations of imidazole). Use HisPur Columns and Cartridges to obtain high-purity His-tagged proteins with no metal contamination.

Highlights:

  • High purity – obtain more than 10 milligrams of pure His-tagged protein per milliliter of resin without optimizing imidazole washing conditions
  • Specificity – cobalt-chelate coordination core binds fewer host protein contaminants, resulting in lower background than nickel resins
  • Low metal leaching – no metal contamination in eluted histidine-tagged protein sample
  • Versatility – purify proteins under native or denaturing conditions; compatible with Thermo Scientific Pierce Cell Lysis Reagents and a variety of buffer additives
  • Flexibility – available as bulk resin, kits, predispensed columns, chromatography cartridges and 96-well filter plates
  • Cost effective – resin can be reused several times or discarded after single use

Available Formats:

Product Details:

Although Ni(2+) chelate resins achieve high protein yields, they bind somewhat indiscriminantly, resulting in suboptimal purity. Often, additional cleanup steps are required. By contrast, cobalt maximizes protein purity without sacrificing protein yield. HisPur Cobalt Resin binds fewer nonspecific proteins, displays less metal leaching and enables less stringent elution conditions than nickel resins.

HisPur Cobalt IMAC Resin specifically binds His-tagged recombinant protein that can be efficiently recovered by mild imidazole elution. Greater than 10mg of His-tagged protein can be bound per milliliter of resin using a 28 kDa fusion protein expressed and purified from an E. coli lysate.

Profile of His-tagged protein purification with HisPur Cobalt Resin
HisPur Cobalt Resin is specific for His-tagged proteins and allows mild, efficient elutions. Cell lysate containing over-expressed recombinant 6xHis-tagged GFP was prepared in B-PER Bacterial Protein Extraction Reagent (Part No. 78243) and protease inhibitors. Protein concentrations were determined by Coomassie Plus Protein Assay (Part No. 23238). Bacterial lysate (1.0mg total protein) was applied to a 0.2mL bed volume of HisPur Cobalt Resin in a spin column. The resin was washed three times with 0.4mL of wash buffer containing 10mM imidazole. His-tagged proteins were eluted three times with 0.2mL of elution buffer containing 150mM imidazole. Gel lanes were normalized to equivalent volume. Gel was stained with Imperial Protein Stain (Part No. 24615). M = Molecular Weight Marker; L = lysate load; FT = flow-through.

Various His-tagged proteins can be recovered in similar or higher purity compared to that obtained from other available nickel or cobalt IMAC resins using either a spin or gravity-flow column format. Purification performance of HisPur Cobalt Resin was benchmarked against IMAC resins from other suppliers using multiple His-tagged proteins of different sizes and expression levels. The results demonstrate that HisPur Cobalt Resin achieves maximal protein purity without sacrificing protein yield.

Highest purity obtained with Thermo Scientific HisPur Cobalt Resin. Comparison of purity of first elution fractions using various Co 2+ and Ni 2+ IMAC Resins. Values were determined by densitometry of stained protein bands in the gels shown in the figure below.
  His-GFP
purity (%)
His-β-Gal
purity (%)
His-Protein L
purity (%)
Thermo Scientific HisPur Cobalt Resin 87 93 77
Supplier C Cobalt Resin 78 90 76
Supplier S Cobalt Resin 85 65 77
Supplier G Nickel Resin 84 83 68
Supplier Q Nickel Resin 85 48 72
Ni +2-IDA Resin 37 16 46

 

Comparison of His-tagged protein purification by HisPur Cobalt Resin and other IMAC cobalt and nickel metal-chelate agarose resin beads.
HisPur Cobalt Resin outperforms other IMAC resins. Comparable yield and higher purity obtained with HisPur Cobalt Resin (lane 1) compared to other IMAC resins (lanes 2 to 6). Cell lysates containing over-expressed recombinant 6xHis-tagged protein were prepared in B-PER Bacterial Protein Extraction Reagent (Part No. 78243) and protease inhibitors. Protein concentrations were determined by Coomassie Plus Protein Assay (Part No. 23238). E. coli lysates containing over-expressed His-tagged GFP, β-galactosidase or Protein L were applied to 0.2mL bed volumes of each IMAC resin in spin column format. Binding, wash and elution buffers were prepared and used per each manufacturer's instructions. The first elution fraction for each IMAC resin was analyzed by SDS-PAGE and protein purity determined by densitometry. Gel lanes were normalized to equivalent volume. Lanes: 1= HisPur Cobalt Resin, 2= supplier C cobalt resin, 3= supplier S cobalt resin, 4= supplier G nickel resin, 5= supplier Q nickel resin, 6= Ni-IDA and L= lysate load.

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
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
89964 HisPur Cobalt Resin
Formulation: Cobalt (IMAC) on crosslinked 6% beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding > 10mg of His-tagged protein per mL of resin
10mL Product Instructions for product #89964 HisPur Cobalt Resin MSDS for product #89964 HisPur Cobalt Resin Certificate of Analysis for product #89964 HisPur Cobalt Resin $92.00

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89965 HisPur Cobalt Resin
Formulation: Cobalt (IMAC) on crosslinked 6% beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding > 10mg of His-tagged protein per mL of resin
100mL Product Instructions for product #89965 HisPur Cobalt Resin MSDS for product #89965 HisPur Cobalt Resin Certificate of Analysis for product #89965 HisPur Cobalt Resin $636.00

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89966 HisPur Cobalt Resin
Formulation: Cobalt (IMAC) on crosslinked 6% beaded agarose, slurried in 20% ethyl alcohol
Sufficient For: Binding > 10mg of His-tagged protein per mL of resin
500mL Product Instructions for product #89966 HisPur Cobalt Resin MSDS for product #89966 HisPur Cobalt Resin Certificate of Analysis for product #89966 HisPur Cobalt Resin $2,510.00

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89967 HisPur Cobalt Spin Columns, 0.2mL
Formulation: 0.8mL-capacity microcentrifuge columns containing 0.2mL resin
Sufficient For: Binding approx. 2mg of His-tagged protein per column
25 columns Product Instructions for product #89967 HisPur Cobalt Spin Columns, 0.2mL MSDS for product #89967 HisPur Cobalt Spin Columns, 0.2mL Certificate of Analysis for product #89967 HisPur Cobalt Spin Columns, 0.2mL $167.00

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89968 HisPur Cobalt Spin Columns, 1mL
Formulation: 8mL-capacity centrifuge columns containing 1mL resin
Sufficient For: Binding approx. 10mg of His-tagged protein per column
5 columns Product Instructions for product #89968 HisPur Cobalt Spin Columns, 1mL MSDS for product #89968 HisPur Cobalt Spin Columns, 1mL Certificate of Analysis for product #89968 HisPur Cobalt Spin Columns, 1mL $110.00

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89969 HisPur Cobalt Spin Columns, 3mL
Formulation: 22mL-capacity centrifuge columns containing 3mL resin
Sufficient For: Binding approx. 30mg of His-tagged protein per column
5 columns Product Instructions for product #89969 HisPur Cobalt Spin Columns, 3mL MSDS for product #89969 HisPur Cobalt Spin Columns, 3mL Certificate of Analysis for product #89969 HisPur Cobalt Spin Columns, 3mL $162.00

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90090 HisPur Cobalt Purification Kit, 0.2mL
Formulation: Multi-component kit
Sufficient For: Purifying approx. 2mg of His-tagged protein per column

Kit Contents:
HisPur Cobalt Spin Columns, 0.2mL, 25 columns
Equilibration/Wash Buffer, 100mL
Elution Buffer, 50mL
Collection Tubes, 80 tubes
25-column kit Product Instructions for product #90090 HisPur Cobalt Purification Kit, 0.2mL MSDS for product #90090 HisPur Cobalt Purification Kit, 0.2mL Certificate of Analysis for product #90090 HisPur Cobalt Purification Kit, 0.2mL $292.00

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90091 HisPur Cobalt Purification Kit, 1mL
Formulation: Multi-component kit
Sufficient For: Purifying approx. 10mg of His-tagged protein per column

Kit Contents:
HisPur Cobalt Spin Columns, 1mL, 5 columns
Equilibration/Wash Buffer, 100mL
Elution Buffer, 50mL
5-column kit Product Instructions for product #90091 HisPur Cobalt Purification Kit, 1mL MSDS for product #90091 HisPur Cobalt Purification Kit, 1mL Certificate of Analysis for product #90091 HisPur Cobalt Purification Kit, 1mL $273.00

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90092 HisPur Cobalt Purification Kit, 3mL
Formulation: Multi-component kit
Sufficient For: Purifying approx. 30mg of His-tagged protein per column

Kit Contents:
HisPur Cobalt Spin Columns, 3mL, 5 columns
Equilibration/Wash Buffer, 225mL
Elution Buffer, 75mL
5-column kit Product Instructions for product #90092 HisPur Cobalt Purification Kit, 3mL MSDS for product #90092 HisPur Cobalt Purification Kit, 3mL Certificate of Analysis for product #90092 HisPur Cobalt Purification Kit, 3mL $292.00

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90093 HisPur Cobalt Chromatography Cartridges, 1mL
Formulation: 1mL resin cartridges with LC and Luer-Lok Fittings
Sufficient For: Binding > 10mg of His-tagged protein per cartridge

Learn more about Pierce Chromatography Cartridges.
5 cartridges Product Instructions for product #90093 HisPur Cobalt Chromatography Cartridges, 1mL MSDS for product #90093 HisPur Cobalt Chromatography Cartridges, 1mL Certificate of Analysis for product #90093 HisPur Cobalt Chromatography Cartridges, 1mL $167.00

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90094 HisPur Cobalt Chromatography Cartridges, 5mL
Formulation: 5mL resin cartridges with LC and Luer-Lok Fittings
Sufficient For: Binding > 50mg of His-tagged protein per cartridge

Learn more about Pierce Chromatography Cartridges.
2 cartridges Product Instructions for product #90094 HisPur Cobalt Chromatography Cartridges, 5mL MSDS for product #90094 HisPur Cobalt Chromatography Cartridges, 5mL Certificate of Analysis for product #90094 HisPur Cobalt Chromatography Cartridges, 5mL $279.00

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90095 HisPur Cobalt Spin Plates
Formulation: 96-well filter plates containing 100┬ÁL HisPur Cobalt 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 #90095 HisPur Cobalt Spin Plates MSDS for product #90095 HisPur Cobalt Spin Plates Certificate of Analysis for product #90095 HisPur Cobalt Spin Plates $334.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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