Home › Biochemistry › Maximizing his-tag protein binding with HisTrap™ excel columns
Steps
  1. 1 Identify the protein binding problem 00:11
  2. 2 Recognize stripping agents as root cause 00:43
  3. 3 Replace conventional IMAC media with HisTrap 00:57
Biochemistry Cytiva

Maximizing his-tag protein binding with HisTrap™ excel columns

Protocol
Difficulty
intermediate

Steps

1
Identify the protein binding problem

The researcher attempts to purify a histidine-tagged protein using conventional HIS-tag columns but experiences poor or no protein recovery despite using the correct methodology.

▶ 00:11
2
Recognize stripping agents as root cause

A colleague explains that chelating agents like EDTA in the sample strip nickel ions from the column resin, preventing his-tag protein binding.

▶ 00:43
3
Replace conventional IMAC media with HisTrap

Switch from standard immobilized metal affinity chromatography (IMAC) columns to HisTrap XL resin from Cytiva to minimize nickel leakage.

▶ 00:57
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