FAQ: What systems does NEB offer for protein expression and purification?

The pMAL™ Protein Fusion and Purification System (NEB# E8200) is available for producing a protein expressed from a cloned gene or open reading frame. The gene of interest is fused to the malE gene, which encodes maltose-binding protein (MBP), and expressed at levels constituting up to 40% of the total cell protein using the tac promoter and translation initiation signals of MBP. A one-step amylose affinity purification for the MBP tag is used to isolate the fusion protein.

The IMPACT™ System (NEB# E6901) utilizes engineered protein splicing elements (inteins) to purify recombinant proteins by a single chitin affinity column. This system distinguishes itself from other protein fusion systems by its ability to separate a recombinant protein from the affinity tag without the use of a protease. In addition, native recombinant proteins possessing a C-terminal thioester can be isolated for applications such as protein semisynthesis and site specific labeling.

The K.lactis Protein Expression Kit (NEB# E1000) provides an easy method for expressing a gene of interest in the yeast Kluyveromyces lactis. Proteins may be produced intracellularly or be secreted using the supplied integrative expression vector pKLAC1.

The PURExpress® In Vitro Protein Synthesis Kit (NEB# E6800) is an E.coli cell-free transcription/translation system reconstituted from purified components. The E.coli transcription/translation factors (except for ribosomes) are His-tagged. Target protein may be expressed from either linear of plasmid DNA templates containing a T7 promoter.

 

The NEBExpress™ Cell-Free E.coli Protein Synthesis System (NEB #E5360) is an extract-based system derived from E. coli cells that have been engineered for high-level in vitro production of recombinant protein from genes cloned downstream of a T7 promoter in either linear or plasmid DNA templates. Efficient isolation of the recombinant target protein may be accomplished by using NEBExpress Ni-NTA Magnetic Beads (NEB #S1423), spin columns (NEB #S1427) or resin (NEB #S1428).

 

NEBExpress™ Ni-NTA Magnetic Beads (NEB #S1423)NEBExpress™ Ni Spin Columns (NEB #S1427) and NEBExpress™ Ni Resin (NEB #S1428) – a portfolio of immobilized Nickel products for the isolation of poly-histidine tagged (His-tagged) proteins from cell lysates or cell-free protein synthesis reactions.