Protein Expression

It’s a matter of expression.

Protein expression can be a very complex, multi-factorial process. Each protein requires a specific intracellular environment to correctly achieve its secondary and tertiary structures. Proteins may also require post-translational modifications or insertion into a cellular membrane for proper function. Other proteins, once expressed, may be toxic to the host. Thus, no single solution exists for the successful production of all recombinant proteins, and a broad range of expression tools is needed to ensure the successful expression of your target protein.

Our NEBExpress portfolio of products includes solutions for expression and purification, and is supported by access to scientists with over 40 years of experience in developing and using recombinant protein technologies in E. coli. We use these solutions in our own research and manufacturing processes, and know that quality and performance are critical – all of our products are stringently tested so that you can be sure they will work optimally for your solution, just as we rely on them to work in ours.

Video: Expression of difficult Proteins

Protein Expression Systems:

Generate analytical amounts of protein with our cell-free expression systems

Generate and purify high yields of recombinant proteins

  • The NEBExpress MBP Fusion and Purification System utilizes a pMAL vector and the malE gene for the expression of MBP-fusion proteins, which can be isolated by a one-step affinity purification
  • The IMPACT Kit utilizes engineered protein splicing elements (inteins) to purify recombinant proteins in a single step
  • For analysis, try our protein standards, which are available unstained, prestained, or with two colors for easy identification

Optimized cells for protein expression

Eukaryotic protein expression

Purify tagged proteins with our magnetic beads, columns and resins

Protein Expression and Purification Selection Chart

Application Kit Advantages
High-Yield Expression pMAL™ Protein Fusion and Purification System Substantial yields (up to 100 mg/L) in more than 75% cases tested; uses the strong Ptac promoter.
K. lactis Protein Expression Kit Uses the strong LAC4 promoter; multiple integrations of plasmid results in higher yield.
IMPACT™ Kit Uses the T7 promoter for high level regulated expression.
Cell-Free Expression PURExpress® In Vitro Protein Synthesis Kit Quickly generates analytical amounts of protein.
Co-Expression of Multiple Proteins K. lactis Protein Expression Kit Easily co-express 2–4 proteins.
PURExpress® In Vitro Protein Synthesis Kit Bicistronic constructs or multiple plasmids with appropriate transcription and translation regulatory elements can be used.
Enhanced Solubility pMAL™ Protein Fusion and Purification System Fusion to MBP enhances solubility of proteins in E. coli.*
K. lactis Protein Expression Kit Utilizes K. lactis eukaryotic folding pathway.
Affinity Tag Chromatography IMPACT™ Kit Utilizes an intein-chitin binding domain (CBD) tag on either the N- or C- terminus, offering single-step purification.
pMAL™ Protein Fusion and Purification System Fusion to MBP allows for purification on amylose resin.
K. lactis Protein Expression Kit Vectors are sold separately that generate fusions to MBP allowing for purification on amylose restin.
Post-translational Modification K. lactis Protein Expression Kit Secretion of both N- and O- glycosylated proteins.
Periplasmic Expression pMAL™ Protein Fusion and Purification System Periplasmic expression enhances folding of proteins with disulfide bonds.
Secreted Expression K. lactis Protein Expression Kit Eliminates cell lysis step, simplifying purification.
Toxic Proteins K. lactis Protein Expression Kit Secretion of protein from the cell.
IMPACT™ Kit Can express the toxic gene in two pieces and ligate proteins together using intein mediated protein ligation (IPL).
pMAL™ Protein Fusion and Purification System Can export toxic proteins into periplasmic space.
PURExpress® In Vitro Protein Synthesis Kit Cell-free environment not affected by “toxicity” of expressed protein.
Protein Modification IMPACT™ Kit Generates proteins with reactive ends (N-terminal cysteine and/or C-terminal thioester) allowing for labeling or ligation of proteins or peptides.
PURExpress® In Vitro Protein Synthesis Kit Allows introduction of modified, unnatural or labeled amino acids.
No Additional Amino Acid Residues IMPACT™ Kit Start of native protein is fused adjacent to site of intein cleavage.
pMAL™ Protein Fusion and Purification System Start of protein is fused adjacent to protease site.

Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.