IDT
IDT
Strep-Tactin® Sepharose®For purification of Strep-tag® II proteins by gravity flow column chromatographyStrep-Tactin® Sepharose® provides high binding capacity and minimal non-specific binding.
The material can be used for gravity flow purification. Since the Strep-Tactin® resins are optimized for column affinity chromatography as opposed to batch purification, we do recommend our pre-packed Strep-Tactin® Sepharose® gravity flow columns, which are available in different formats (from 0.2 ml mini columns up to 10 ml columns).
For packing individual columns bulk material is available.
In one step: over 99% purity and high purification factors A 36 kDa enzyme and a mutant, both with C-terminal Strep-tag® II, were expressed in the cytoplasm of E. coli. The crude lysate was chromatographically separated on Strep-Tactin® Sepharose (5 mg/ml) under gravity flow and physiological conditions (100 mM Tris-Cl, pH 8.0). The purification is documented on a Coomassie stained SDS gel where samples from the crude lysate (lane 1), from the flow through (lane 2), and from the elution with 2.5 mM desthiobiotin (lane 3) had been applied. The wild type enzyme is shown to be over 99 % pure (lane 3 contains 50 µg protein vs 0.5 µg protein per band in the molecular size standard). The mutant - although expressed at low level only - could be obtained at high purity under the same conditions. 1: Sample of crude lysate after cytosolic expression with pASK-IBA3 2: Sample of flow through during chromatography 3: Sample after the addition of 2.5 mM desthiobiotin M: Molecular size standard (kDa) Specifications of Strep-Tactin® Sepharose® resin
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