pcDNA™6 BioEase™ Gateway® Biotinylation System

Catalog number: K98001

Invitrogen™  Related applications: Gateway Cloning | Protein Expression

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Description

The pcDNA™6 BioEase™ Gateway® Biotinylation System provides an easy, efficient method for expressing, purifying, and detecting biotinylated recombinant proteins. The pcDNA™6/BioEase™-DEST vector (Figure 1) incorporates a 72 amino acid sequence from K. pneumoniae that directs in vivo biotinylation of a specific lysine residue. This K. pneumoniae tag is recognized and efficiently biotinylated in vivo by a native mammalian system similar to the enzyme Biotin Protein Ligase (BPL) in E. coli(1). Proteins produced from the pcDNA™6/BioEase™-DEST vector are expressed as fusion proteins to this sequence. Biotinylated fusion proteins can be purified by streptavidin agarose, detected on western blots using streptavidin-HRP or streptavidin-AP conjugates, or visualized in mammalian cells by immunofluorescence with a streptavidin-FITC conjugate (Figure 2). In addition to the 72 amino acid K. pneumoniae sequence, the pcDNA™6/BioEase™-DEST vector includes the following features:

attR sites for fast and efficient recombination with any attL-flanked Gateway® entry vector
• The CMV promoter for high-level expression of the recombinant fusion protein
• An enterokinase cleavage site for efficient removal of the N-terminal BioEase™ fusion tag
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: CMV
Protein Tag or Fusion: BioEase Tag
Delivery Method: Transfection
Cloning Method: Gateway®
Selection Agent (Eukaryotic): Blasticidin
Vector Type: pDEST, pcDNA
Cleavage: EK (Enterokinase) Recognition Site
Constitutive or Inducible System: Constitutive
Product Size: 20 reactions

Contents & storage

The pcDNA™6/BioEase™-DEST Gateway® Biotinylation System includes the pcDNA™6/BioEase™-DEST vector, pcDNA™6/BioEase™-GW/lacZ control vector, and Streptavidin-HRP conjugate. Store at -20°C. All components are guaranteed stable for 6 months when properly stored.

Documents

Manuals & protocols