HisPur Cobalt Resin, Columns and Purification Kits
Specific, fast and gentle 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.
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
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.
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.
Thermo Scientific HisPur Cobalt Resin
Supplier C Cobalt Resin
Supplier S Cobalt Resin
Supplier G Nickel Resin
Supplier Q Nickel Resin
Ni +2-IDA Resin
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.