Tips for winning chess against His-tagged proteins

Purification of recombinant proteins fused to a polyhistidine tag by immobilized metal affinity chromatography (IMAC) has become routine in many labs. But do you use the best fitting matrix in every case to enable efficient purification? Surprisingly, when asked, most researchers cannot explain why they choose one resin over another for their His-tagged protein. Read the expert’s opinion before setting up your next experiment.

NTA and IDA ligands, loaded with nickel or cobalt, are the most commonly used ones in commercially available resins for His-tagged protein purification. But which one is better; when and why? The detailed overview for a right case-specific resin selection “NTA vs. IDA: a tale of two ligands” from Cube Biotech protein purification specialists will help you to navigate and to make the best choice depending on your protein needs.

Cube Biotech experts always advise: “Let your protein determine your ligand and metal ion and do not forget to consider what else is in your sample. When deciding on the best IMAC resin for your protein purification needs, first answer a few questions:

  • How pure does your target protein need to be?
  • What yield of target protein do you need?
  • What yields do you anticipate from your expression?
  • Does your sample contain reagents like DTT or EDTA?
  • What are your budgetary constraints?

Theory is not enough; own experience rules your choices. Try free samples of best in class IMAC resins from Cube Biotech available at flash4science for USA, UK and German scientists this week:

Image  by chrisgj6  / CC BY-SA 2.0

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