An affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins in manual or automated formats. Immobilized Metal Affinity Chromatography (IMAC) purifications employing Ni-NTA (nickel-nitrilotriacetic acid) magnetic beads can be performed under native or denaturing conditions which permit efficient binding and purification of insoluble proteins, proteins that aggregate in inclusion bodies, or proteins that possess a tertiary structure that sequester the polyhistidine affinity tag. Immobilized His-tagged proteins can be used in subsequent experiments to pull-down proteins that may interact with the immobilized protein.
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