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GE28-9545-52

pGEX-4T-3

Cytiva 28-9545-52

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About This Item

UNSPSC Code:
41106501
NACRES:
NA.56

packaging

pkg of 25 μg

manufacturer/tradename

Cytiva 28-9545-52

shipped in

dry ice

storage temp.

−20°C

General description

Thirteen pGEX vectors are available (see Figure). Nine of the vectors have an expanded multiple cloning site (MCS) that contains six restriction sites. The expanded MCS facilitates the unidirectional cloning of cDNA inserts obtained from libraries constructed using many available lambda vectors. pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 each encode the recognition sequence for site-specific cleavage by PreScission Protease, (see PreScission Protease) between the GST domain and the multiple cloning site. pGEX-4T-1, pGEX-4T-2, and pGEX-4T-3 are derived from pGEX-2T and contain a Thrombin recognition site. pGEX-5X-1, pGEX-5X-2, and pGEX5X-3 are derivatives of pGEX-3X and possess a Factor Xa recognition site.

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

Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Legal Information

pGEX Vectors
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.

This page shows how to cleave and purify GST-tagged proteins bound to Glutathione Sepharose in batch mode from Cytiva. Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, and Glutathione Sepharose 4B can all be used for cleavage and purification of GST-tagged proteins in batch.

This page shows how to cleave and purify GST-tagged proteins eluted from GSTrap from Cytiva.

This page shows how to cleave and purify GST-tagged proteins bound to GSTrap from Cytiva.

Protocols

Preparing Cells for Electroporation

This page describes preparation of competent cells and transformation with pGEX DNA.

This page describes the characteristics of pGEX expression vectors used with Cytiva products.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service