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  • Video: Capturem his maxiprep
  • Video: Capturem his miniprep
  • Visual protocol: Capturem his maxiprep
  • Visual protocol: Capturem his miniprep
  • Capturem nickel column reagent compatibility
  • TALON reagent compatibility
  • His60 reagent compatibility
  • TALON: Native vs denaturing purification
  • Protocol: denaturing purification with TALON resin, imidazole elution
  • Protocol: native purification with TALON resin, imidazole elution
  • Protocol: native purification with TALON resin, pH elution
Selection guides His60 selection guide
Home › Learning centers › Protein research › His-tag purification › Protocols › His60 reagent compatibility

His-tag purification

  • Purification methods overview
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  • Selection guide: His60 resin
  • xTractor Buffer is optimized for superior protein yield
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  • Tech note: cobalt resin
  • Simplified purification of active, secreted his-tagged proteins
  • Overview: His60
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    • Video: Capturem his maxiprep
    • Video: Capturem his miniprep
    • Visual protocol: Capturem his maxiprep
    • Visual protocol: Capturem his miniprep
    • Capturem nickel column reagent compatibility
    • TALON reagent compatibility
    • His60 reagent compatibility
    • TALON: Native vs denaturing purification
    • Protocol: denaturing purification with TALON resin, imidazole elution
    • Protocol: native purification with TALON resin, imidazole elution
    • Protocol: native purification with TALON resin, pH elution
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Selection guides His60 selection guide

His60 reagent compatibility

Overview

His60 Ni resin is a high-capacity Ni-IDA resin that permits one-step protein purification of his-tagged proteins under either native or denaturing conditions. The resin is compatible with multiple denaturants and detergents—refer to the User Manual for a complete reagent compatibility table. Purified recombinant his-tagged proteins can be eluted by the addition of 300 mM imidazole or by a reduction in pH.

Reagents compatible with His60 Ni resins
ReagentAcceptable Concentration
Beta-mercaptoethanola 20 mM (with caution)
CaCl2 5 mM
CHAPSb,c 1% (with caution)
DTT (dithiothreitol)d 1 mM (with caution)
Ethanole 20%
Glycerol 20%
Guanidine hydrochloridef 6 M
HEPESg Up to 100 mM (with caution)
Histidineh
  • 20 mM to inhibit nonspecific binding
  • Up to 100 mM to elute his-tagged proteins
MgCl2 4 M
MOPSg Up to 100 mM (with caution)
NP-40b,i 2%
SDSb,c 1% (with caution)
Sodium acetate Up to 100 mM (with caution)
Sodium phosphate Up to 50 mM
Trisj Up to 50 mM (with caution)
Triton-X 100b,i 1%
Tween 20 2%
Ureaf 8 M
  1. Use the resin immediately after equilibrating with buffers containing beta-mercaptoethanol. Otherwise, a slight change in color (yellowing of the resin) will occur. Do not store the resin in buffers containing beta-mercaptoethanol.
  2. Detergents cannot be easily removed by buffer exchange.
  3. Ionic detergents like CHAPS (3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate), SDS (sodium dodecyl sulfate), and sarkosyl are compatible up to 1%. However, due to their charged nature, you should anticipate interference with binding, even at low concentrations.
  4. Since DTT is a reducing agent, low concentrations will reduce the metal ions in His60 Ni Superflow resin. Although enough of these ions may remain unaffected to allow protein purification, please use it with caution. Perform at least 20 column volumes of washes, preferably with low concentrations of imidazole (40 mM) to wash out any reduced metal ions.
  5. Ethanol may precipitate proteins, causing low yields and column clogging.
  6. With high concentrations, protein unfolding generally takes place. Protein refolding on-column (or after elution) is protein-dependent.
  7. Amine groups that are present in these buffers can interact with Ni2+ ions, diminishing the resin’s binding capacity.
  8. Binds to His60 Ni and competes with histidine residues in the his-tag.
  9. Has high absorbance at 280 nm.
  10. Tris coordinates weakly with metal ions, causing a decrease in capacity.
Reagents incompatible with His60 Ni Superflow Resin
These reagents are incompatible at any concentration:
  • Arginine, glycine, and glutamine
  • EDTA (ethylenediaminetetraacetic acid), EGTA (ethylene glycol tetraacetic acid), and PEI
    NOTE: Using these chelating agents will strip metal ions from the resin, resulting in protein elution and a resin color change.
  • DTE (dithioerythritol)

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  • Viral detection with qPCR
  • SARS-CoV-2 pseudovirus
  • Human ACE2 stable cell line
  • Viral RNA isolation
  • Viral and host sequencing
  • Vaccine development
  • CRISPR screening
  • Drug discovery
  • Immune profiling
  • Publications
  • Next-generation sequencing
  • Spatial omics
  • RNA-seq
  • DNA-seq
  • Single-cell NGS automation
  • Reproductive health
  • Bioinformatics tools
  • Immune profiling
  • Real-time PCR
  • Great value master mixes
  • Signature enzymes
  • High-throughput real-time PCR solutions
  • Detection assays
  • References, standards, and buffers
  • Stem cell research
  • Media, differentiation kits, and matrices
  • Stem cells and stem cell-derived cells
  • mRNA and cDNA synthesis
  • In vitro transcription
  • cDNA synthesis kits
  • Reverse transcriptases
  • RACE kits
  • Purified cDNA & genomic DNA
  • Purified total RNA and mRNA
  • PCR
  • Most popular polymerases
  • High-yield PCR
  • High-fidelity PCR
  • GC rich PCR
  • PCR master mixes
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  • In-Fusion seamless cloning
  • Competent cells
  • Ligation kits
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  • Automated platforms
  • Plasmid purification kits
  • Genomic DNA purification kits
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