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Strep-Tactin Superflow Plus

For batch, gravity-flow, or FPLC purification of Strep-tagged proteins
  • 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
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 coupled to magnetic beads for small-scale purification procedures (up to 100 μg) or 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.

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Cat No./ID: 30004
Strep-Tactin Superflow Plus (10 ml)
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For purification of Strep-tagged proteins: 10 ml Strep-Tactin-charged Superflow Plus suspension (50%) containing 0.02% sodium azide (max. pressure: 140 psi)
Cat No./ID: 30020
Strep-Tactin Superflow Plus Cartridge (1 ml)
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1 cartridge pre-filled with 1 ml Strep-Tactin Superflow Plus (contains 0.02% sodium azide): for automated purification of Strep-tagged proteins using liquid chromatography systems
Cat No./ID: 30060
Strep-Tactin Superflow Plus Cartridge (5 ml)
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1 cartridge pre-filled with 5 ml Strep-Tactin Superflow Plus (contains 0.02% sodium azide): for automated purification of Strep-tagged proteins using liquid chromatography systems
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.

Product Details

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Highly specific detection of Strep-tagged proteins in any expression system.
Purified Strep-tagged thioredoxin (10 ng) was mixed with crude cell lysate from mammalian (NIH-3T3), insect (Sf9), and bacterial (E. coli) cells (each 10 µg total protein). After electrophoresis and western transfer, the Strep-tagged protein was detected using the Strep-tag Antibody and chemiluminescent detection.
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Efficient purification of Strep-tagged GFP.

GFP was expressed in 60 ml E. coli culture and a 5 ml aliquot was applied to Strep-Tactin Superflow Plus. Protein was eluted using 2.5 mM desthiobiotin, giving a total protein yield of 3.1mg. C: Cleared lysate; E: Elution fractions; F: Flow-through; M: Markers; W: Wash.
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Binding of Strep-tagged proteins to Strep-Tactin and elution using desthiobiotin.
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

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 PlusUp to 9 mg/ml
Strep-Tactin SuperflowUp to 3 mg/ml (~150 nmol@ 20 kDa)
Strep-Tactin Magnetic Beads200-300 μg/ml (10% suspension) (~10-15 nmol @ 20 kDa)

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 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.

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

Specifications

Features
Specifications
Applications Proteomics
Binding capacity up to 9 mg/ml
N- or C-terminal tag Both
Processing manual/automated
Scale Strep-Tactin Superflow Plus: Large Scale; Strep-Tactin Superflow Plus Cartridge: medium scale
Start material cell lysates
Support/matrix Superflow Plus
Tag Strep-tag
Yield up to 9 mg/ml
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