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packaging
pkg of 25 μg
manufacturer/tradename
Cytiva 28-9546-48
shipped in
dry ice
storage temp.
−20°C
General description
pGEX-2TK is designed to allow the detection of expressed proteins by directly labeling the fusion products in vitro (1). This vector contains the recognition sequence for the catalytic subunit of cAMP-dependent protein kinase obtained from heart muscle. The protein kinase site is located between the GST domain and the MCS. Expressed proteins can be directly labeled using protein kinase and [gamma-32P]ATP and readily detected using standard radiometric or autoradiographic techniques. pGEX-2TK is a derivative of pGEX-2T; its fusion proteins can be cleaved with Thrombin.
Cleavage of pGEX-6P GST fusion proteins occurs between the Gln and Gly residues of the recognition sequence Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro (2). Low temperature (5°C) digestion minimizes the degradation of the protein of interest. Because PreScission Protease has been engineered with a GST tag, it can also be removed from the cleavage mixture simultaneously with the GST portion of the fusion protein. The pGEX-6P Expression Vectors permit convenient site-specific cleavage and simultaneous purification on Glutathione Sepharose. The pGEX-6P series provides all three translational reading frames linked between the GST coding region and the multiple cloning site.
Collectively, the pGEX vectors provide all three translational reading frames beginning with the EcoRI restriction site. pGEX-1λT, pGEX-6P-1, pGEX-4T-1, and pGEX-5X-1 can directly accept and express cDNA inserts isolated from λgt11 libraries.
Features and Benefits
- A tac promoter for chemically inducible, high-level expression of GST-tagged recombinant proteins.
- An internal lacIq gene for use in any E. coli host.
- Very mild elution conditions for release of fusion proteins from the affinity matrix, thus minimizing effects on antigenicity and functional activity.
- PreScission Protease, Thrombin, or Factor Xa recognition sites for cleaving the desired protein from the fusion product.
Storage and Stability
Analysis Note
Legal Information
pGEX Vectors are to be used for scientific investigation and research and for no other purpose whatsoever and a license for commercial use of the licensed products and the processes claimed in US patent 5,654,176 and equivalent patents and patent applications in other countries must be negotiated directly with Millipore Corp (formerly Chemicon International Inc) by the purchaser prior to such use.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Certificates of Analysis (COA)
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Articles
This page describes troubleshooting strategies for cloning the gene or gene fragment into a pGEX expression vector using GE Healthcare products.
This page shows how to cleave and purify GST-tagged proteins bound to Glutathione Sepharose in batch mode from GE Healthcare.
This page shows how to cleave and purify GST-tagged proteins bound to GSTrap from GE Healthcare.
This page shows how to cleave and purify GST-tagged proteins eluted from GSTrap from GE Healthcare.
Protocols
This page describes ligation of insert to pGEX DNA using GE Healthcare products.
This page describes electroporation using GE Healthcare products.
This page describes preparation of competent cells and transformation with pGEX DNA.
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