Technology
The Strep-tag story
Principle & properties

Cloning/gene expression

Purification
Introduction
Applications
Resin Specifications
Strep-Tactin Sepharose
Strep-Tactin Superflow
Strep-Tactin MacroPrep
Strep-Tactin POROS
MagStrep
Column formats
Spin columns
High-throughput
 purification
Antibody columns
Buffers & Reagents
Strep-tag Starter Kits

Detection
Introduction
Strep-Tactin HRP
Strep-Tactin AP
Antibodies
Fluorescence detection
Protein ladder

Diagnostic/Assay/
Immobilization

Examples

References

FAQ

Specifications of Strep-Tactin resins Reagents compatible with the Strep-tag/Strep-Tactin interaction Five Strep-Tactin resin versions are available which differ in their properties and applications. While Strep-Tactin Sepharose® is preferentially used for gravity flow chromatography, Strep-Tactin® Superflow®, Strep-Tactin® Superflow® high capacity and Strep-Tactin MacroPrep® can also be used for low pressure or FPLC applications, and Strep-Tactin POROS® (20 and 50) for FPLC and HPLC chromatography. Each matrix exhibits different non-specific binding properties. In cases where unsatisfactory results are obtained with one matrix due to unspecific background, it is advisable to switch to one of the other matrices (see also Strep-Tactin Column Evaluation Sets). In addition, Superflow is especially suited for increased flow rates and is also useful for the purification of large protein complexes. Please note, that the Strep-Tactin resins are optimized for column affinity chromatography as opposed to batch purification. Therefore, pre-packed columns with different Strep-Tactin resins are available. For batch purification use our MagStrep Strep-Tactin coated Magnetic Beads. In table 2, reagents compatible with the Strep-tag/Strep-Tactin interaction are listed. Table 1: Specifications of Strep-Tactin resins Strep-Tactin Sepharose Strep-Tactin Superflow Strep-Tactin Superflow high capacity Strep-Tactin MacroPrep Strep-Tactin POROS 20 Strep-Tactin POROS 50 MagStrep Magnetic Beads Gravity flow column Yes Yes Yes Yes No No No Batch purification For One- Strep-tag only For One- Strep-tag only For One- Strep-tag only For One- Strep-tag only For One- Strep-tag only For One- Strep-tag only Yes** FPLC No Yes Yes Yes Yes Yes No HPLC No Yes Yes Yes Yes Yes No Binding capacity 50 - 100 nmol/ml 50 - 100 nmol/ml 150 - 500 nmol/ml 50 - 100 nmol/ml 25 - 50 nmol/ml 25 - 50 nmol/ml Type1: 100pmol/mg beads Type2: 300pmol/mg beads Support Sepharose 4FF Superflow 6 Superflow 6 MacroPrep 50 POROS 20 POROS 50 / Bead structure agarose 6% agarose, crosslinked 6% agarose, crosslinked polymethacrylate polystyrene polystyrene magnetite content Type1: 50-60 % Type2: 20-40% Bead size 45 - 165 µm 60 - 160 µm spherical 60 - 160 µm spherical 50 µm 20 µm 50 µm Type1: <0.5-1.2 µm Type2: 0.5-1.5 µm Exclusion limit 3 x 107 6 x 106 6 x 106 1 x 106 n.d. n.d. / Recommended linear flow rate* up to 30 cm/h up to 300 cm/h up to 300 cm/h up to 300 cm/h 300 - 500 cm/h 300 - 500 cm/h / pH stability 4 - 9 2 - 13 2 - 13 2 - 12 1 - 13 1 - 13 n.d. Extreme pH as listed above may be used for short cleaning procedures but should be changed to pH 8.0 for storage and Strep-tag protein binding Max. pressure Gravity flow n.d n.d n.d 170 bar 170 bar / Form buffered 50% suspension buffered 50% suspension buffered 50% suspension buffered 50% suspension buffered 50% suspension buffered 50% suspension 50 mg/ml suspension in Dulbecco´s PBS; 0.1 % bovine serum albumin; 0.02 % NaN3 Storage 4 °C, do not freeze 4 °C, do not freeze 4 °C, do not freeze 4 °C, do not freeze 4 °C, do not freeze 4 °C, do not freeze 4 °C, do not freeze Shipment RT RT RT RT RT RT RT nd = not determined; *linear flow rate calculation ** For Strep-tagII only recommended if expression rate >1mg/ml otherwise use One-Strep-tag Strep-Tactin Column Evaluation Set: Strep-Tactin MacroPrep, Superflow and Sepharose (left to right). Scale: reduced. Strep-Tactin columns regenerated with HABA (see also Strep-tag purification cycle): Strep-Tactin MacroPrep, Superflow and Sepharose (left to right). Since MacroPrep is not as transparent as Sepharose or Superflow, the color shift to red is less visible. Scale: reduced. Table 2: Reagents compatible with the Strep-tag/Strep-Tactin interaction Reagent Concentration Reduction Agents DTT 50 mM β-mercaptoethanol 50 mM Non-Ionic Detergents C8E4 Octyltetraoxyethylene max. 0.88 % C10E5; Decylpentaoxyethylene 0.12 % C10 E6 0.03 % C12 E8 0.005 % C12E9; Dodecyl nonaoxyethylene (Thesit) 0.023 % DM; Decyl-β-D-maltoside 0.35 % LM; N-dodecyl β-D-maltoside 0.007 % NG; N-nonyl-β-D-glucopyranoside 0.2 % OG; N-octyl-β-D-glucopyranoside 2.34 % TX; Triton X-100 2 % Tween 20 2 % Ionic Detergents N-lauryl-sarcosine 2 % 8-HESO;N-octyl-2-hydroxy-ethylsulfoxide 1.32 % SDS; Sodium-N-dodecyl sulfate 0.1 % Zwitter-Ionic Detergents CHAPS 0.1 % DDAO; N-decyl-N,N-dimethylamine-N-oxide 0.034 % LDAO; N-dodecyl-N,N-dimethylamine-N-oxide 0.13 % Others Ammonium sulfate (NH4)2SO4 2 M CaCl2 max. 1 M EDTA 50 mM Ethanol 10 % Guanidine max. 1 M Glycerol max. 25 % Imidazole max. 250 mM MgCl2 1 M NaCl 5 M Urea max. 1 M Note: These reagents have been successfully tested for the purification of e.g. GAPDH-Streptag with concentrations up to those mentioned. For reagents not marked with "max." higher concentrations may be possible, though. Since binding depends on the sterical accessibility of Strep-tag in the context of the particular protein the possible concentration may deviate from the given value for other proteins. Protocols for download

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