Spin columns containing an affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins. Immobilized Metal Affinity Chromatography (IMAC) purifications employing NEBExpress® Ni Spin columns can be performed under native or denaturing conditions, yielding highly pure target in a single protein purification step. This enables screening of expression conditions and streamlines the functional and structural characterization of the target protein.
Support Matrix: Spherical, agarose based microparticles ranging in size from 10-100 μm.
Figure 1: NEBExpress Ni Spin Column Quick Start ProtocolFigure 2: Purification of His-tagged fusion proteins using NEBExpress Ni Spin Columns NEBExpress Ni Spin Columns were equilibrated and loaded with 500 μl of crude extract from E. coli containing a plasmid that expresses either His6-GluRS or His6-NDK fusion protein. Columns were washed, eluted and analyzed by SDS-PAGE to confirm low non-specific binding of extract proteins and high isolation of target. Marker (M) contains Color Prestained Protein Standard (NEB #P7719), Lane 1: GluRS protein load, Lane 2: GluRS flow through, Lane 3: GluRS 1st elution, Lane 4: GluRS 2nd elution, Lane 5: NDK protein Load, Lane 6: NDK flow through, Lane 7: NDK 1st elution, Lane 8: NDK 2nd elution.
This product is related to the following categories:
Amylose Purification (MBP-tag),
Nickel Purification (His-tag),
Affinity Purification,
Protein Purification
This product can be used in the following applications:
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