Efficiently express your target protein by cloning your gene into one of our panel of carefully designed expression plasmids. Offering a choice of bacterial, mammalian or insect cell systems, EMD Millipore’s vast portfolio of expression vectors enables you to choose the perfect combination of promoters, epitope tags, antibiotic resistance, and host compatibility. With our vectors and cloning kits, you can generate high performance constructs in as little as 40 minutes.
pET E. coli T7 Expression Vectors
The pET System is the most powerful system for the cloning and expression of recombinant proteins in E. coli. Driven by the strong bacteriophage T7 promoter and translation signals, Novagen’s® pET System has been used to express thousands of different proteins in host cells expressing T7 polymerase. The pET System has continuously expanded to offer new technologies and options for expression, and includes a large collection of pET vector types, different host strains and companion products designed for efficient detection and purification of target proteins.
- pET-31b(+) for high yield bioproduction of peptides and small proteins
- pET-32a-c for production of soluble, active target proteins in E. coli
- pET-39b and 40b using Dsb tags for export and periplasmic folding of target proteins
- pET-41a-c and 42a-c with popular GST fusion tags for enhanced production and solubility
- pET-43.1a-c designed for cloning and high-level expression of polypeptide sequences fused with the 495 aa NusA (Nus•Tag™) protein
- pET-44a-c with Nus•Tag sequence plus N- and C-terminal His•Tag sequences
- pET-45b(+) with amino-terminal His•Tag™ sequence and minimal extraneous sequences
- pET-46 Ek/LIC prepared vector for ligation-independent cloning, with amino-terminal His•Tag sequence
- pET-47b(+) through pET-50b(+) with HRV 3C Protease cleavage site for efficient fusion tag removal
Nus•Tag™ fusion increases the solubility of recombinant annexin A1.
pENTR™ vectors containing the annexin A1 target gene were generated using the Gateway Nova pET-53-DEST™ vector (lanes 2-3, pET-His•Tag®/annexin-A1/Strep•Tag® II) and Gateway Nova pET-57-DEST vector (lanes 4-5, pET-His•Tag/Nus•Tag/thrombin/annexin-A1/Strep•Tag II).
Soluble annexin A1 was purified on sequential His•Bind and Strep•Tactin columns (lanes 6-7). The N-terminal fusion tags were removed by thrombin cleavage and a subtractive His•Bind column (lanes 8-9).
Proteinase K
Use Proteinase K to stop enzymatic reactions, including endonucleolytic cleavage and polynucleotide addition, and efficiently remove proteins from nucleic acid preparations. Proteinase K is a highly active serine protease that exhibits broad cleavage specificity on native and denatured proteins and is widely used in the purification of DNA and RNA. Its activity is increased in the presence of denaturants such as SDS (1%) and elevated temperature (50-60°C). The enzyme is available lyophilized or as a ready-to-use concentrated stock solution (600 mAU/mL). Novagen Proteinase K products are free of detectable DNase and RNase.
