Ni Sepharose™ excel Medien für die immobilisierte Metallionen-Affinitätschromatographie (IMAC)
Lieferant: Cytiva
Ni Sepharose™ excel ist ein mit sehr stark an einen chelatbildenden Liganden gebundenen Nickelionen vorgeladenes IMAC-Medium. Proben, die normalerweise ein Stripping von Metallionenverursachen, können daher auf das Medium geladen werden. Ni Sepharose™ excel ist hauptsächlich für das Einfangen und Reinigen von Histidin-markierten Proteinen, die in Zellkulturüberstände aus eukaryotischen Zellen, wie Insektenzellen oder CHO-Zellen, abgesondert werden.
- Load eukaryotic cell culture samples containing secreted histidine-tagged proteins directly with retained binding capacity
- Increase target protein yield and decrease degradation through reduced and simplified sample handling
- Choose between several formats for screening and preparative purification of histidine-tagged proteins
Ni Sepharose excel enables direct loading of samples without having to perform extensive and time-consuming pretreatment procedures. The flow properties of the medium make it excellent for purifications in a wide range of scales and allow loading of large sample volumes, enabling purification of low concentrations of target proteins at large volumes. Dynamic binding capacity was tested with 0,5 mg/ml (histidine)6-tagged pure protein (Mr 43 000) in EX-CELL™ 420 Insect serum-free medium (capacity at 10% breakthrough) or (histidine)6-tagged protein (Mr 28 000). Column volume was 1 ml and flow rate 1 ml/min. Binding capacity is sample-dependent.
Samples that usually cause stripping of metal ions can therefore be loaded to the medium. The nickel ions have been shown to remain bound to the chelating ligand even after incubation for 24 hours in 10 mM EDTA. Ni Sepharose excel is designed primarily for capture and purification of histidine-tagged proteins secreted into cell culture supernatants from eukaryotic cells such as insect cells or CHO cells.
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