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Thermo Scientific™ HisPur™ Ni-NTA Spin Purification Kit, 3 mL

Catalog No. PI88229
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5 Column Kit
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PI88229 5 Column Kit
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Catalog No. PI88229 Supplier Thermo Scientific™ Supplier No. 88229
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Purify His-tagged recombinant fusion proteins using high-capacity nickel-NTA (nitrilotriacetic acid) metal-chelate beads, spin columns and plates.

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

The specially prepared nickel-NTA 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 and chromatography cartridges

  • Cost-effective – reuse the same batch of resin at least five times

  • Easy to use – pre-formulated buffers available for kit formats

Available Formats:

  • Resin Slurries – crosslinked 6% beaded agarose; 10mL, 100mL, 500mL bottles
  • Spin Columns – 0.2mL (microcentrifuge), 1mL, 3mL columns
  • Purification Kits – complete kits in all three column sizes
  • Chromatography Cartridges – compatible with automated liquid chromatography systems and with syringe processing
  • Bulk and Custom – let us know your needs for bulk HisPur Ni-NTA Resin

Includes:

Kits contain HisPur Ni-NTA Columns

Specifications

Capacity (Metric) 1 mL/Column
Description HisPur Ni-NTA Spin Purification Kit, 3mL
Content And Storage Sufficient For: Purifying up to 180 mg of His-tagged protein per column
• HisPur Ni-NTA Spin Columns, 3 mL, 5 columns
• Phosphate-buffered Saline (10X), 30 mL
• Imidazole (2M), 25 mL
• Column Plugs, 5 plugs

Store at 4°C.
Product Line HisPur
Quantity 5 Column Kit
Target His-Tagged Protein
Type Spin Column Kit
Stationary Phase Divalent Nickel, Metal Chelate
Sufficient For 180 mg of His-Tagged Protein per Column
Format Spin Column
Column Type Affinity, Agarose Resin
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Are there other sequences that can bind to nickel more tightly than 6xHis-tagged proteins and how can they be eluted.

Yes, 7xHis-tagged proteins, proteins naturally high in histidine, and other combinations of His and other amino acids will bind. To elute them, you have to increase the concentration of imidazole. Generally these peptides will not contaminate your fraction since they remain on the column. However after multiple uses of the same column, these peptides may reduce the binding capacity of the column.

Can ProBond or Ni-NTA beads be used for large-scale preparations?

ProBond and Ni-NTA beads can be used in FPLC columns. However, the beads can only withstand low pressure (~43.5 psi max).

What should the typical protein recovery be when using the Probond Purification System or Ni-NTA Purification system?

Both systems are qualified by purifying 2 mg of myoglobin protein on a column and performing a Bradford assay. Protein recovery must be 75% or higher.

What is the difference between HisPur Nickel Resin and HisPur Cobalt Resin?

Nickel has a higher affinity for histidines than does cobalt. It binds multiple histidines more tightly than cobalt, and requires more stringent conditions than is necessary for cobalt. The lower affinity of cobalt for multiple histidines typically results in less nonspecific binding of histidine-rich proteins that lack a his-tag compared to nickel resins.

HisPur Ni-NTA resin has a high capacity of up to 60 mg of 6xHis-tagged protein per milliliter. It is a versatile resin that can work under both native and denaturing conditions, and can also be used with a variety of lysis reagents and buffer additives.

HisPur Cobalt resin utilizes proprietary tetradentate chelating resin charged with cobalt. This system recovers highly purified protein with lower imidazole concentrations and has low metal leeching properties.

For Research Use Only. Not for use in diagnostic procedures.