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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

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Catalog number selected: V2020

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PinPoint™ Xa Protein Purification System
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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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Download SDSPDF (225 KB) – English (United States)

Certificate of Analysis

See all certificates for V2020

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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Manas

India