His Tag Antibodies

Structure of His Tag

His Tag is a kind of polypeptide tag with a relatively small molecular weight, usually composed of 6 to 10 histidine residues, and the molecular weight is approximately 0.8 to 1.2 kDa. The molecular weight of this tag varies slightly among different variants, depending on the amount of histidine and the design of the connection sequence.

His Tag forms stable complexes by coordinating and binding with metal ions such as nickel and cobalt through its continuous histidine residues. After the tag is fused and expressed at the N-terminal or C-terminal of the protein, it usually does not significantly affect the folding and function of the target protein. Its binding mechanism depends on the interaction between the nitrogen atom on the histidine imidazole ring and the immobilized metal ion, and can achieve efficient purification under denatured or non-denatured conditions. This tag is colorless, has low immunogenicity, and is easily eluted through EDTA or imidazole competition. It has become one of the most commonly used tools in the purification of recombinant proteins.

Fig. 1 His-Tag-induced higher-order oligomers.¹

Key structural properties of His Tag:

• Short peptide composed of continuous histidine residues linear structure
• Histidine imidazole ring nitrogen atoms as metal ligand bonding sites
• Fuse to the N-terminal or C-terminal of the target protein through genetic engineering

Functions of His Tag

The main function of His Tag is to serve as an affinity tag in the expression and purification of recombinant proteins. In addition, it also plays a key role in a variety of biotechnology applications, including protein detection, immobilization and functional research.

The binding of His Tag with metal ions has high affinity and reversibility. It can maintain the binding ability under different pH and denaturation conditions (such as high-concentration urea or guanidine hydrochloride), and is suitable for the renaturation purification of denatured proteins. Its binding ability increases with the increase of histidine number. The 6×His tag achieves the optimal balance between binding strength and tag size and has become one of the most commonly used affinity tags in industrial production and scientific research.

Applications of His Tag and His Tag Antibody in Literature

1. Bromberg, Raquel, et al. "The His-tag as a decoy modulating preferred orientation in cryoEM." Frontiers in Molecular Biosciences 9 (2022): 912072. https://doi.org/10.3389/fmolb.2022.912072

This paper finds that the presence of His-tag affects the interaction between proteins and the gas-liquid interface during cryo-electron microscopy sample preparation and alters their dominant orientation distribution. Studies have shown that His tag can mask the hydrophobic/hydrophilic regions on the protein surface, and its hydrophobic linker may also enhance the interfacial interaction, suggesting that tag design can regulate protein-interface interactions.

2. Ayoub, Nooraldeen, et al. "Structural and biochemical insights into His-tag-induced higher-order oligomerization of membrane proteins by cryo-EM and size exclusion chromatography." Journal of Structural Biology 215.1 (2023): 107924. https://doi.org/10.1016/j.jsb.2022.107924

This study takes two His-tagged membrane proteins as examples and finds that His tags can mediate the formation of higher-order oligomers (such as tetramers, decamers, etc.). Chromatographic and cryo-electron microscopy results revealed tag-mediated specific interactions, indicating that ignoring tag removal may interfere with downstream protein structure and function studies.

3. de Almeida, Janaina Marques, et al. "Tailoring recombinant lipases: keeping the His-tag favors esterification reactions, removing it favors hydrolysis reactions." Scientific reports 8.1 (2018): 10000. https://doi.org/10.1038/s41598-018-27579-8

This study explored the influence of His tags on the lipase LipC12. After removing the His tag, the enzymatic hydrolytic activity was significantly enhanced, but the activity and stability of the enzyme in organic solvents were better when the tag was present. The His tag does not affect the immobilization efficiency, but it has different effects on the catalytic activity of free and immobilized enzymes, indicating that the tag can be retained or removed according to application requirements.

4. Jureczek, Justyna, et al. "An oligo-His-tag of a targeting module does not influence its biodistribution and the retargeting capabilities of UniCAR T cells." Scientific Reports 9.1 (2019): 10547. https://doi.org/10.1038/s41598-019-47044-4

This study explored the influence of oligo-His tags of targeted modules (TM) in UniCAR systems on their functions and distribution. By comparing the anti-PSCA TM with and without His tags, it was found that the His tag had no significant effect on its binding affinity, in vitro and in vivo killing effect, and biological distribution.

5. Mišković, Marija Zora, et al. "Location is everything: influence of his-tag fusion site on properties of adenylosuccinate synthetase from Helicobacter pylori." International journal of molecular sciences 25.14 (2024): 7613. https://doi.org/10.3390/ijms25147613

This study took the adenylate succinate synthase (AdSS) of Helicobacter pylori as an example and found that the N-terminal His tag would cause protein aggregation and precipitation, significantly reducing the enzyme activity, while the C-terminal tag had little effect. The results show that the position of the His tag has a crucial impact on protein solubility and function, and careful evaluation is required when making a selection.

Creative Biolabs: His Tag Antibodies for Research

Creative Biolabs specializes in the production of high-quality His Tag antibodies for research and industrial applications. Our portfolio includes monoclonal antibodies tailored for ELISA, Flow Cytometry, Western blot, immunohistochemistry, and other diagnostic methodologies.

• Custom His Tag Antibody Development: Tailor-made solutions to meet specific research requirements.
• Bulk Production: Large-scale antibody manufacturing for industry partners.
• Technical Support: Expert consultation for protocol optimization and troubleshooting.
• Aliquoting Services: Conveniently sized aliquots for long-term storage and consistent experimental outcomes.

For more details on our His Tag antibodies, custom preparations, or technical support, contact us at email.