Purification of His-Tagged Proteins using Ni-NTA Beads v2

• DOI: 10.17504/protocols.io.kqdg3wz8ev25/v2
• Publication Date: 2025-05-30T12:29:51Z
• Authors: Mariángeles Ávila...; Antonella Denise ...; Karla Vanessa Gaona; María Teresa Damiani

This protocol describes the batch purification of recombinant His-tagged proteins expressed in E. coli. Proteins containing a 6xHis tag are purified using Ni-NTA (Nickel-Nitrilotriacetic Acid) beads. The His-tag allows selective binding to nickel ions, enabling efficient purification under native conditions. After cell lysis, the lysate is incubated with Ni-NTA beads, followed by washing and elution using imidazole.