Gene Expression & Protein Purification Products, NEB

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Home > Technical Reference > Gene Expression & Protein Purification

Gene Expression & Protein Purification Products

Protein expression and purification can be very challenging because there is no single approach that is appropriate for every protein or every downstream application. Successful expression can be affected by each individual protein's intrinsic ability to fold, its solubility, its toxicity or its need for post-translational modifications. Additionally, the application for which the protein is ultimately to be used may dictate how the protein needs to be expressed or purified. It is therefore important to have a flexible set of tools and methods that can be applied to each unique expression and purification project.
Each NEB expression technology offers different advantages which enable you to choose the strategy that best suits your protein expression and purification needs. Various applications along with the recommended NEB kit are highlighted below.

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 may result in higher yield.
	IMPACT™ Kit	Uses the T7 promoter for higher expression and tight control.
Enhanced solubility	pMAL™ Protein Fusion and Purification System	Fusion to MBP enhances solubility of proteins in E. coli.*
Enhanced solubility	K. lactis Protein Expression Kit	Utilizes K. lactis eukaryotic folding pathway.
Affinity tag chromatography	IMPACT™ Kit	Utilizes an intein-CBD tag on either the N- or C- terminus.
Affinity tag chromatography	pMAL™ Protein Fusion and Purification System	Fusion to MBP allows for purification on amylose resin.
Post-translational modification	K. lactis Protein Expression Kit	Secretion of both N- and O- glycosylated proteins.
Post-translational modification	RheoSwitch® Mammalian Inducible Expression System	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, simplifying purification.
Secreted Expression	RheoSwitch® Mammalian Inducible Expression System	Eliminates cell lysis, simplifying purification.
Characterization of eukaryotic proteins in homologous systems	RheoSwitch® Mammalian Inducible Expression System	Synthetic inducer and engineered receptor eliminate non-specific side effects.
Toxic genes	K. lactis Protein Expression Kit	Utilizes a LAC4 promoter that has been modified to lack background expression in E. coli.
	RheoSwitch® Mammalian Inducible Expression System	Precise regulation of expression.
	IMPACT™ Kit	Can express the toxic gene in two pieces and ligate proteins together.
Protein labeling or ligation	IMPACT™ Kit	Elutes proteins with reactive ends (N-terminal cysteine and/or C-terminal thioester). The C-terminal thioester can be labeled after purification.
No additional amino acid residues	IMPACT™ Kit	Native protein sequence is fused adjacent to site of cleavage.

* Kapust and Waugh (1999) Protein Science, 8, 1668–1674.