Hexa-His  (Synonyms: 6X His Tag)

Hexa-His (6X His Tag) is a commonly used affinity tag made up of six histidine residues. HEXA-HIS can bind to affinity chromatography media containing transition metal ions like nickel (Ni²⁺) or cobalt (Co²⁺), making it useful for protein purification^[1][2].

Hexa-His for Protein Purification and Protein Analysis^[2]
(1) Construct a plasmid containing the target gene and Hexa-His tag, and introduce it into E. coli.
(2) Select positive clones and grow them in LB medium until the OD600 is about 0.6-0.8, then add IPTG (HY-15921, isopropyl β-D-thiogalactoside) as an inducer and continue to incubate for a few hours to induce the expression of the target protein.
(3) Collect the bacterial cells, resuspend them in lysis buffer, and sonicate to lyse the cells, then centrifuge to remove debris and obtain the supernatant.
(4) Prepare the Ni-NTA affinity chromatography column and slowly pass the lysate supernatant through the Ni-NTA column to bind the CTB protein with the Hexa-His tag to the column.
(5) After protein adsorption, wash the resin with 10 column volumes of 60 mM imidazole binding buffer. Elute the target gene with the same solution containing 1 M imidazole using 5 column volumes. Collect the protein and analyze it by 15% SDS-PAGE.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

840.85

C₃₆H₄₄N₁₈O₇

64134-30-1

Solid

White to off-white

His-His-His-His-His-His

HHHHHH

Room temperature in continental US; may vary elsewhere.

Sealed storage, away from moisture

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

• [1]. Watly J, et al. Insight into the coordination and the binding sites of Cu(2+) by the histidyl-6-tag using experimental and computational tools. Inorg Chem. 2014 Jul 7;53(13):6675-83.  [Content Brief]

  [2]. Ana Paula de Mattos Arêas, et al. Synthesis of cholera toxin B subunit gene: cloning and expression of a functional 6XHis-tagged protein in Escherichia coli.Protein Expr Purif.2002 Aug;25(3):481-7.  [Content Brief]