Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

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Purification of His-tagged Proteins

Ready-to-Use Ni-IDA Columns – First Choice for Higher Yields of His-tagged Proteins

MobiSpin Ni-IDA Columns

MobiSpin Ni-IDA Columns

IDA-Structure

Silica-Based Ni-IDA Matrix
The Ni-IDA silica is an affinity chromatography matrix for purifying recombinant proteins carrying a His-tag. Histidine residues in the His-tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions which enables strong and efficient binding of target protein. In free Ni-IDA the vacant positions are usually occupied by water molecules (as shown above).

Ni-IDA Purification Procedure

Purification Procedure
The purification of His-tagged proteins consists of 4 steps: cell lysis, loading & binding, washing, and elution. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted with buffer from the matrix.

Features

  • Columns are pre-packed and ready-to-use
  • Highest flexibility of applications – protein purification by gravity flow or by spinning
  • Easy, fast, and cost-effective routine purification of recombinant polyhistidine-tagged proteins
  • Purification under native and denaturating conditions
  • Starting from diverse expression systems, e.g., E. coli, yeast, insect, and
    mammalian cells
  • Universal use – suitable for small proteins, large protein complexes, proteins with low expression rates
  • High binding capacity and high affinity
  • Improved target specificity by optimized silica-based Ni2+-IDA matrix
  • Imidazol free loading and washing buffer
  • Columns are long-term storable when kept dry
  • Simply replace your current Ni-NTA products, no optimization or protocol change necessary!

Description

  • Ni-IDA Columns provide a fast and convenient routine tool for purification of recombinant polyhistidine-tagged proteins.
  • The form-stable silica matrix is precharged with Ni2+ ions and allows purification on the principle of Immobilized Metal Ion Affinity Chromatography (IMAC). Binding of proteins is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni2+ ions. The chelating group of the Ni-IDA resin is based on IDA (iminodiacetic acid), which enables strong and efficient binding of target protein onto the IMAC matrix.
  • In contrast to traditional IDA matrices, MoBiTec Ni-IDA is an optimized matrix with low density of IDA ligands. This non-saturating surface concentration of IDA eliminates almost all non-specific interactions of contaminating host proteins with the adsorbent. As a result, MoBiTec Ni-IDA provides higher target protein purity.
  • IDA is a tridentate chelator which occupies three of the six binding sites in the coordination sphere of the Ni2+ ion. The remaining three coordination sites are usually occupied by water molecules and can be exchanged with histidine residues of the recombinant protein.

MobiSpin Ni-IDA Columns

MobiSpin Ni-IDA Columns

MobiSpin Ni-IDA Columns

Features

  • For protein purification by spinning
  • Very high binding capacity: up to 12 mg protein per spin column
  • Excellent protein recovery rate of > 90%
  • Simultaneous processing of multiple samples

MoBiTec Ni-IDA Columns

MoBiTec Ni-IDA Columns

MoBiTec Ni-IDA Columns

MoBiTec Ni-IDA Columns with bound His6-GFP

MoBiTec Ni-IDA Columns
Equilibrated with Lysis-Equilibration-Wash Buffer (left) and with bound His6-GFP (right).

Features

  • For protein purification by gravity flow
  • Maximal binding capacity: up to 90 mg protein per column
  • Protein recovery rate of > 80%
  • Columns are long-term storable and reusable

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
PR-HTK105 MobiSpin Ni-IDA Columns 5 columns 69,00 PDF
PR-HTK110 MobiSpin Ni-IDA Columns 10 columns 103,00 PDF
PR-HTK004 MoBiTec Ni-IDA Columns 4 columns 108,00 PDF
PR-HTK010 MoBiTec Ni-IDA Columns 10 columns 244,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

Related Products

Order# Description Amount Price Data Sheet
TOPO-HIS01 His6-GFP bacterial expression vector 10 µg 459,00 PDF
PEG01 pEG-His 1 vector 5 µg 242,00 PDF
PR-SB01-01 SpeedBlot (His) 30 ml 214,00 PDF
HIST01 anti-HIS-Tag antibody 100 µl 277,00 PDF
D291-11 Anti-His-tag mAb-Magnetic Beads 20 Tests (Slurry: 1 mL) 270,00 PDF
D291-10 Anti-His-tag mAb-Magnetic Agarose 20 Tests (Gel: 200 μl) 297,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.
  • D291-11 and D291-10 are only available in Germany; distributed for MBL.

Download

  • Purification of His-tagged Proteins Flyer (PDF)
  • SpeedBlot (His) Western Blot Stain Flyer (PDF)