For general questions please contact our Customer Service:
EMD Millipore
290 Concord Road
01821 Billerica, MA
United States
Phone: 781 533-6000
Fax: 800-645-5439
E-mail: Customer.CareDesk@emdchemicals.com
25 May 2013
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This product has been discontinued.
pET Expression Systems and pET Expression Systems plus Competent Cells provide core reagents needed for target gene cloning and expression.
Components: pET Expression Systems
Components for pET Expression Systems are similar for all systems unless otherwise stated with the specific pET Expression System description. pET Expression Systems include:
• 10 µg pET vector DNA (for each indicated plasmid)
• 0.2 ml BL21 Glycerol Stock
• 0.2 ml BL21(DE3) Glycerol Stock
• 0.2 ml BL21(DE3)pLysS Glycerol Stock
• 0.2 ml Induction Control Glycerol Stock
Components: pET Expression Systems plus Competent Cells
pET Expression Systems plus Competent Cells contain all of the components of the specific pET Expression System, as well as the following additional components, unless otherwise stated with the specific pET Expression System description:
• 0.2 ml NovaBlue Competent Cells
• 0.2 ml BL21(DE3) Competent Cells
• 0.2 ml BL21(DE3)pLysS Competent Cells
• 2 × 0.2 ml SOC Medium
• 10 µl Test Plasmid
These components are sufficient for up to 10 transformations in each host.
Purification and Detection Reagents
Purification and detection reagents are available separately. For complete product descriptions and ordering information, refer to the Protein Purification and Antibodies, Conjugates & Detection Tools chapters.
pET Expression System 49b
The pET-49b(+) vector is designed for cloning and high-level expression of target proteins fused with the 220 aa schistosomal glutathione-S-transferase (GST•Tag™) protein that is cleavable with the human rhinovirus (HRV) 3C protease (1). Schistosomal glutathione-S-transferase (GST) is commonly used as a fusion partner when expressing proteins in E. coli (2). The GST•Tag sequence has been reported to enhance the production and in some cases the solubility of its fusion partners. When expressed in a soluble, properly folded form, GST•Tag fusion proteins can be purified with immobilized glutathione. Gentle elution is achieved with buffers containing reduced glutathione. Quantification of soluble GST fusions is also possible by assaying the glutathione-S-transferase activity. Cloning sites are available for producing fusion proteins also containing a cleavable N- terminal His•Tag® sequence for detection and purification. The plasmid contains a strong T7lac promoter, optimized RBS, the coding sequence for the HRV 3C protease cleavage site (LeuGluValLeuPheGlnGlyPro) for N-terminal fusion tag removal and a multiple cloning site that contains restriction enzyme sites found in many other Novagen expression vectors to facilitate insert transfer. An optional C-terminal S•Tag™ coding sequence is compatible with purification, detection, and quantification (3).
The pET System allows for expression of target genes under control of the T7 promoter and is covered by US Patent 5,693,489. This patent is licensed by EMD Millipore Corporation from Brookhaven National Labs. Commercial entities need to obtain a license from Brookhaven National Labs prior to use of this Product. Ccommercial use of this Product also requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
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| Fusion tag | His•Tag, GST•Tag, S•Tag | |||||||||||||||||
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| Storage | ≤ -70°C | ||
| Ship |
Shipped with Blue Ice or with Dry Ice
Standard Handling |
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