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PinPoint™ Xa Protein Purification System

Produce and Purify In Vivo Biotinylated Fusion Proteins

  • In vivo biotin tags make purification easy; many proteins produced are soluble
  • Purify fusion proteins by column or batch methods
  • PinPoint™ Vectors are supplied for all reading frames

Size

Catalog number selected: V2020

$ 708.00 Your price: Log In

PinPoint™ Xa Protein Purification System
1 system
$ 708.00
Your price: Log In

Express and Purify In Vivo-Biotinylated Fusion Proteins

Express and purify fusion proteins that are biotinylated in vivo. The DNA coding for the protein of interest is cloned into a PinPoint™ Vector downstream of a sequence encoding a peptide that becomes biotinylated in vivo. Biotinylated fusion proteins are produced in E. coli and are affinity-purified using the SoftLink™ Soft Release Avidin Resin. This proprietary resin allows elution of the fusion protein under nondenaturing conditions. The PinPoint™ Vectors feature the encoded endoproteinase Factor Xa (pronounced "ten a") proteolytic site that provides a way to separate the purification tag from the native protein, and the vectors carry a convenient multiple cloning region for ease in construction of fusion proteins.

The system contains vectors in all possible sense reading frames, an avidin-conjugated resin, Streptavidin-Alkaline Phosphatase, a purification column and biotin. The PinPoint™ Xa Control Vector contains the chloramphenicol acetyltransferase (CAT) gene provided as a means of monitoring protein expression, purification and processing conditions. The system generally yields 1–5mg of protein per liter of culture.

Additional features include the flexibility to use SoftLink™ Resin for release of the fusion protein under nondenaturing conditions and presence of the tac promoter, which allows tightly regulated expression.

Applications

  • Purification of milligram amounts of expressed protein.
  • Production of novel bioreagents.

Reference

  1. Cress, D. et al. (1993) Promega Notes 42, 2–7.
0923VA-W

PinPoint™ Xa-1 Vector map.

Sequence Information

PinPoint™ Xa-1 Vector GenBank® Accession Number U47626.

PinPoint™ Xa-2 Vector GenBank® Accession Number U47627

PinPoint™ Xa-3 Vector GenBank® Accession Number U47628.

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

SoftLink™ Soft Release Avidin Resin

V201C 1 × 3ml

PinPoint™ Xa-1 Vector

V203A 1 × 10μg View Product

PinPoint™ Xa Control Vector

V204A 1 × 5μg

PinPoint™ Xa-2 Vector

V205A 1 × 10μg

PinPoint™ Xa-3 Vector

V206A 1 × 10μg

PinPoint™ Purification Column

V556A 1 × 1 each

Biotin

V557A 1 × 1ml100mM

Streptavidin Alkaline Phosphatase

V559A 1 × 20μl View Product

SDS

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

CC

See Protocol for detailed storage recommendations.

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