It is often desirable to design a method of purification directly into a recombinant protein expression strategy. This is most often accomplished by expressing a target protein with an appended “tag” that permits its purification by affinity chromatography. Tags can be a short peptide that specifically interacts with an immobilized antibody or metal ion, or larger binding domains that interact with specific immobilized ligands.
An important consideration in using a protein-tagging strategy is whether or not the presence of a tag will affect the protein’s downstream applications. For example, the presence of a larger binding domain may permit more simple purification, but it might also adversely affect the biochemical function of the target protein it is fused to. Therefore, it is often desirable to include a method for tag removal following purification of the fusion protein. Common methods include the use of site-specific proteases to cleave at an engineered site between the tag and the target protein, or the use of an intein to auto-catalyze tag removal.
- Co-expression of Multiple Proteins in Kluyveromyces lactis
- Intein-Mediated Protein Ligation (IPL) and Labeling with the IMPACT™ Kit
- Protein Expression with T7 Express Strains
- Transformation of SHuffle® Competent Cell Strains
- Use of the PURExpress® in vitro Protein Synthesis Kit, Disulfide Bond Enhancer and SHuffle® Competent E. coli for heterologous in vitro and in vivo cellulase expression.
The Future of Cell-Free Protein Synthesis
Cell-free protein synthesis has the potential to become one of the most important high throughput technologies for functional genomics and proteomics.
Why Choose the K. lactis Protein Expression Kit?
Review the advantages of the K. lactis Protein Expression Kit for rapid, high yield protein expression in yeast.
Avoid Common Obstacles in Protein Expression
Read how to avoid common obstacles in protein expression that prevent interactions with cellular machinery.
Protein Expression & Purification Brochure
The Protein Expression and Purification brochure provides information on the advanced tools for protein expression and purification offered by NEB.
- IMPACT™ Vectors and Applications
- Protein Expression and Purification Selection Chart
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.