cOmplete™ His-Tag Purification Resin and Columns from Roche

Protein Protection with Immobilized Metal Ion Affinity Chromatography

Immobilized Metal Ion Affinity Chromatography (IMAC) using nickel is a method for protein purification, but it’s toxicity level may affect the protein and increase costs of toxic waste disposal. The presence of nickel can adversely affect the yield and function of your target protein, as metal ions are known to accelerate oxidation of proteins. Common proteases identify oxidative modifications as markers and prefer degradation of these proteins.

cOmplete™ His-Tag Purification Resin and Columns from Roche are proven to have the tightest nickel ion binding, even under harsh conditions, keeping your protein safe beyond DTT and EDTA, and increasing your lab safety. cOmplete™ His-Tag Purification Resin features nickel-chelate chemistry that minimizes nickel ion leakage, tightly binding the nickel ion to the sepharose bead with more links than any conventional IMAC resin.

  • Choose buffer conditions that suit your protein. Use the concentrations your protein requires without loss of capacity or purity, even under harsh conditions (see Figures 1 and 2).
  • Obtain highly pure and less aggregated protein. Minimize nickel leakage to avoid protein oxidation and aggregation (see Figure 3).
  • Avoid toxic nickel with your protein and in your lab. Reuse the resin multiple times without recharging nickel ions (see Figures 1 and 2).
  • Use the prepacked columns with your FPLC system. cOmplete™ His-Tag Purification Columns are fully compatible with the most commonly used purification systems.

Tight Binding

Figure 1: Tight binding of nickel. Before (left) and after (right) photos after 5 times reuse without recharging. Result: The resin remains blue, indicating that there is little to no nickel leakage.

 

Protein-binding performance with His6 CFP.

Figure 2: Protein-binding performance with His6 CFP. cOmplete™ His-Tag Purification Resin (blue columns) with 10 mM DTT and EDTA is reused without nickel recharging alongside Resin G (grey columns) with 1 mM EDTA and 5 mM DTT (as specified in manufacturer’s package insert). Another competing product, Resin Q (not shown), did not bind any protein at all.

Loss of resin Ni ions under stringent conditions

Figure 3: Loss of resin Ni ions under stringent conditions. One milliliter each of cOmplete™ His-Tag Purification Resin and 2 commercially available resins were incubated in 9 ml of a buffer containing 50 mM NaH2PO4, 300 mM NaCl, pH 8.0, 10 mM EDTA, 10 mM dTT, and 500 mM imidazole. Result: The cOmplete™ His-Tag Purification Resin lost less than 1% of nickel ions.

Materials