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11438 - DM Issue 3 - v3 8/2/06 1:14 pm Page 14

Solutions for histidine-tagged protein purification
Immobilized Metal Affinity Chromatography (IMAC) is an Both Ni Sepharose and IMAC Sepharose offer:
excellent chromatography technique for purification of
histidine-tagged proteins1. Pure protein can be obtained > Stability in a wide range of denaturants, detergents,
from a cell extract in a single purification step. The and buffer systems
(histidine)6-tag has become the most used affinity tag
> Compatibility with commonly used reducing agents,
due to its small size, strong metal ion binding, and ability
such as DTT, DTE, TCEP, and β-mercaptoethanol
to bind under denaturing conditions. IMAC with Ni2+ is the
standard method for purifying histidine-tagged proteins, > Higher protein binding capacity than that found with
but in some cases other metal ions give the best results. comparable media from other manufacturers
GE Healthcare offers four media for IMAC in a variety of > Low metal ion leakage
formats for efficient purification of histidine-tagged
proteins. Ni Sepharose™ High Performance and Ni Use the selection guides below to choose the
Sepharose 6 Fast Flow are precharged with Ni2+, while best product to meet your purification needs.
IMAC High Performance and IMAC Sepharose 6 Fast
Flow are uncharged and can be used with a metal ion
of your choice. See licensing information on page 28.

11438 - DM Issue 3 - v3 8/2/06 1:14 pm Page 15

Fig 1. Purifying histidine-tagged proteins with His GraviTrap™ is
a simple four-stage procedure using gravity flow.

Fig 2. His MultiTrap™ HP plates and His MultiTrap FF are 96-well

filter plates that deliver high well-to-well (shown above) and plate-
to-plate reproducibility for screening large numbers of samples.

Fig 3. His SpinTrap™ columns are designed for efficient, small-scale

purification of histidine-tagged proteins directly from clarified or
unclarified cell lysates. The columns are used with a standard

Fig 4. HiTrap™ IMAC HP can be charged with the metal ion of your
choice (see article on page 22).