Qiagen, 30004, Strep-Tactin Superflow Plus (10 ml)

For purification of Strep-tagged proteins: 10 ml Strep-Tactin-charged Superflow Plus suspension (50%) containing 0.02% sodium azide (max. pressure: 140 psi)

Features

- Three-fold increase in binding capacity (up to 9 mg/ml)
- Ideal for purification from prokaryotic and eukaryotic systems
- Enables automated purification on any liquid chromatography system
- Minimal influence on protein structure, function, and immunogenicity

Product Details

The small Strep- tag II (WSHPQFEK) can be used to purify active, native conformation proteins to high purity on Strep- Tactin resins. In addition, it can be used in combination with the His tag in a standardized two-step purification procedure. Strep- Tactin is available as Superflow chromatography resin for larger scale purification (up to 9 mg/ml resin). Strep- Tactin resin is also available in prefilled cartridges for automated purification on liquid chromatography systems.

Performance

Strep- Tactin Superflow Plus allows highly efficient purification of proteins (see figures Efficient purification of Strep-tagged GFP with Strep-Tactin Superflow Plus and Highly specific detection of Strep-tagged proteins in any expression system ).

Meanwhile, pQE TriSystem His· Strep TriSystem Vectors can be used to express proteins doubly-tagged with a His- and a Strep -tag, which are purified in a two-step affinity procedure. Sequential Ni-NTA and Strep -Tactin purification steps are standardized, very fast, and deliver ultrapure proteins.

Principle

Matrix: Protein binding capacity
Strep -Tactin Superflow Plus: Up to 9 mg/ml

Strep- tagged proteins bind with high affinity and specificity to Strep- Tactin, an engineered form of streptavidin (see table Protein binding capacities of Strep- Tactin matrices). After washing, proteins are eluted using low concentrations of biotin (the natural ligand of streptavidin) or desthiobiotin (a stable, reversibly binding biotin analog, see figure Binding of Strep-tagged proteins to Strep-Tactin and elution using desthiobiotin ). Labeling proteins with the small Strep- tag II enables efficient, standardized purification of recombinant proteins expressed in prokaryotic and eukaryotic systems, and delivers highly pure proteins. In addition, the Strep- tag II is the system of choice in situations where other tags may be unsuitable, e.g., when buffer additives such as EDTA may strip IMAC columns.

Procedure

Nonionic detergents: Ionic and zwitterionic detergents
2% Triton X-100: 2% N-lauryl sarcosine
2% Tween 20: 0.1% SDS
2% N-octyl-β-D-glucopyranoside: 1.3% N-octyl-2-hydroxy-ethylsulfoxide
0.2% N-nonyl-β-D-glucopyranoside: 0.3% CHAPS
2% Igepol CA-630 (Nonidet P40): 0.1% N-dodecyl-N,N-dimethylamine-N-oxide
0.12% C10E5; decylpentaoxyethylene: 0.034% DDAO;N-decyl-N,N-dimethylamine-N-oxide
0.03% C10 E6: 10% ethanol
0.005% C12 E8
0.023% C12E9; dodecyl nonooxyethylene (Thesit)
0.35% DM; decyl-β-D-maltoside
0.007% LM; N-dodecyl-β-D-maltoside

Strep -tagged proteins are expressed using the pQE TriSystem Vector and obtained at very high purity from cell lysates using Strep -Tactin Superflow Plus affinity purification procedures. Strep -tagged proteins can be purified using batch or gravity-flow methods, or FPLC-compatible columns can be filled with Strep -Tactin Superflow Plus for purification on liquid chromatography systems. Strep -Tactin Superflow Plus is available in 5 ml cartridges for automated purification on liquid chromatography systems such as the FPLC, ÄkTA, and BioLogic systems, or manual purification using a syringe. For reagents compatible with the Strep- Tactin resins, see table Reagents compatible with the Strep- tag Strep- Tactin interaction.

The C-terminal location of the tag ensures that only full-length proteins are purified. The purification process is highly reproducible, guaranteeing the same high quality protein preparations time after time. Monoclonal Strep- tag antibodies are used to detect Strep -tagged proteins in immunodetection procedures with high sensitivity and specificity.

Applications

- Functional or interaction studies
- Crystallization screening
- Structural analysis
- Generation of antibodies

Strep -Tactin Superflow Plus can be used for efficient and reproducible purification of up to 9 mg Strep -tagged protein per ml resin. Purified proteins can be used for: