PrepEase Histidine-tagged Protein Purification Midi Kit - High Specificity  

Binding Capacity up to 2.5 mg
Culture Volume up to 150 ml

Designed for fast, convenient and high specificity purification of His-tagged proteins

• High Specificity - due to single binding site to His-tagged protein
• Low metal leaching - 5 binding sites to Ni²⁺
• High Purity > 95%
• Includes PrepEase High Specificity His-Tag Protein Columns and all the necessary reagents for easy protein purification
• Room temperature storage

PrepEase® His-Tagged Protein Purification Mini Kit - High Specificity
Binding Capacity up to 400 μg
Culture Volume up to 25 ml

PrepEase® His-Tagged Protein Purification Maxi Kit - High Specificity
Binding Capacity up to 5.0 mg
Culture Volume up to 300 ml

The PrepEase Protein Purification Kits for High Specificity are designed to purify recombinant polyhistidine-tagged (His-tagged) recombinant proteins expressed in E. coli using immobilized metal ion affinity chromatography (IMAC). The kits may also be used to purify His-tagged proteins from other expression systems such as insect cells, mammalian cells, and yeast. Purification may take place under standard, native conditions or under denaturing conditions depending on the solubility of the expressed protein.

PrepEase Protein Purification Kits are based on the use of novel, dry silicabased resin, precharged with Ni²⁺ ions. The High Specificity Kits utilize TED (tris-carboxymethyl ethylene diamine), a special chelating group, which enables strong and efficient binding of target proteins to the resin. TED is a strong pentadentate metal chelator, which occupies five of the six binding sites in the coordination sphere of the Ni²⁺ ion. The single remaining binding site can be exchanged with the histidine residues of the recombinant protein.

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Binding of His-tagged Protein to PrepEase Ni-TED Resin

Purification methods that utilize other chelators, such as NTA (nitrilotriacetic acid), may lead to metal leaching and/or non-specific binding of untagged proteins to the matrix. NTA chelating groups occupy four of the six binding sites    in the coordination sphere of the Ni²⁺ ion, leaving two sites for protein binding. The additional chelating site of TED minimizes metal leaching during purification and reduces non-specific binding of untagged proteins to the matrix. This results in high specificity of His-tagged protein binding to the resin, leading to higher purity of the eluted target protein.

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PrepEase Silica-Based Resin Compared to Agarose Beads

PrepEase Protein Purification Kits

• Novel - Based on dry silica based resin
• Less non-specific binding of contaminating proteins compared to Ni agarose beads
• Easy to elute protein off column
• Utilizes metal (Ni) affinity gravity-flow chromatography
• High stability against chelating and reducing agents
• Room temperature storage