Strep-Tactin Superflow Plus

For batch, gravity-flow, or FPLC purification of Strep-tagged proteins

S_1926_ADNA_PROT_113

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Strep-Tactin Superflow Plus (10 ml)

Cat. No. / ID:  30004

For purification of Strep-tagged proteins: 10 ml Strep-Tactin-charged Superflow Plus suspension (50%) containing 0.02% sodium azide (max. pressure: 140 psi)
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SuspensionCartridge
Superflow Plus
Superflow Plus Cartridge
The Strep-Tactin Superflow Plus is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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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.

See figures

Principle

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
See figures

Procedure

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.

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.

Applications

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:

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

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsProteomics
Binding capacityup to 9 mg/ml
ScaleStrep-Tactin Superflow Plus: Large Scale; Strep-Tactin Superflow Plus Cartridge: medium scale
TagStrep-tag
Support/matrixSuperflow Plus
Yieldup to 9 mg/ml
Processingmanual/automated
Start materialcell lysates
N- or C-terminal tagBoth

FAQ

How often can the Strep-Tactin Superflow Plus resin be reused?
The Strep-Tactin Superflow Plus resin can be reused up to 5 times.
FAQ ID -2169
Are there any changes in the procedure using the Strep-Tactin Superflow Plus resin with the higher capacity over the standard matrix?

No, the protocols are unchanged. The higher binding capacity of Strep-Tactin Superflow Plus resin results in elution fractions with higher protein concentration, compared to standard Strep-Tactin Superflow.

 

 

 

FAQ ID -2168