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Cat. No. / ID: 30004
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✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
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.
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.
| Matrix | Protein binding capacity |
|---|---|
| Strep-Tactin Superflow Plus | Up to 9 mg/ml |
| Strep-Tactin Superflow | Up to 3 mg/ml (~150 nmol@ 20 kDa) |
| Strep-Tactin Magnetic Beads | 200-300 μg/ml (10% suspension) (~10-15 nmol @ 20 kDa) |
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 pre-filled 1 ml cartridges and 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.
| Nonionic detergents | Ionic and zwitterionic detergents | Reducing agents | Others | Salts |
|---|---|---|---|---|
| 2% Triton X-100 | 2% N-lauryl sarcosine | 50 mM DTT | 25% glycerol | 5 M sodium chloride |
| 2% Tween 20 | 0.1% SDS | 2 M ammonium chloride | ||
| 2% N-octyl-β-D-glucopyranoside | 1.3% N-octyl-2-hydroxy-ethylsulfoxide | 1 M calcium chloride | ||
| 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 | 1 M guanidine | ||
| 0.12% C10E5; decylpentaoxyethylene | 0.034% DDAO;N-decyl-N,N-dimethylamine-N-oxide | 1 mM phenylmethyl-sulfonyl fluoride (PMSF) | ||
| 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 |
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.
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:
| Features | Specifications |
| Applications | Proteomics |
| Binding capacity | up to 9 mg/ml |
| Scale | Strep-Tactin Superflow Plus: Large Scale; Strep-Tactin Superflow Plus Cartridge: medium scale |
| Tag | Strep-tag |
| Support/matrix | Superflow Plus |
| Yield | up to 9 mg/ml |
| Processing | manual/automated |
| Start material | cell lysates |
| N- or C-terminal tag | Both |
