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The ready-to-use GoBio™ Mini NiMAC column comprises prepacked WorkBeads™ NiMAC resin which is pre-charged with strongly bound nickel ions, providing very high resistance to reducing agents such as DTT and also to the chelator EDTA. The strong nickel ion binding of this resin is key to its suitability for purification of large volumes of His-tagged proteins from various sources, such as eukaryotic cell extracts, and it requires minimal sample pre-treatment that would normally cause metal ion stripping.
GoBio Mini NiMAC are available in two column sizes: 1 mL and 5 mL for quick and convenient purification of His-tagged proteins by using Immobilized Metal Ion Affinity Chromatography (IMAC).
GoBio Mini NiMAC columns can be used to purify up to 50 mg or 250 mg protein using a 1 mL or a 5 mL column.
- Ready-to-use columns for fast results
- Resin with extra strongly bound Ni2+ resulting in extremely low nickel ion leakage
- Highly resistant to reducing agents up to 20 mM DTT
- Highly resistant to chelating substances present in eukaryotic extracts or up to 20 mM EDTA
- High purity and reproducible results
Table 1. Main characteristics of WorkBeads NiMAC resin.
| WorkBeads NiMAC | |
| Target substances | His-tagged proteins |
| Matrix | Highly cross-linked agarose |
| Average particle size1 (Dv50) | 45 µm |
| Precharged ions | Nickel (ll) ions, Ni²⁺ |
| Static binding capacity |
> 80 mg/mL resin |
| Dynamic binding capacity² |
> 40 mg/mL resin |
| Metal ion capacity³ |
> 60 μmol Cu²⁺/mL resin |
|
Max flow rate (20 cm bed height and 5 bar)⁴ |
600 cm/h |
| Chemical stability |
Compatible with all standard aqueous buffers used for protein purification, and additives such as 20 mM NarEDTA, 20 mM dithiothreitol (DTT), 20 mM TCEP, 20 mM ß-mercaptoethanol, 8 M urea, 6 M guanidine-HCI, non-ionic detergents, 500 mM imidazole, 30% isopropanol, 0.5 M NaOH |
| pH stability |
3–9 (working range) 2–14 (cleaning-in-place) |
| Storage | 2 to 25 °C in 20% ethanol |
¹ The median particle size of the cumulative volume distribution.
² Binding capacity may vary depending on protein characteristics and on flow rate used. A lower flow rate usually increases the dynamic binding capacity.
³ Metal ion capacity is determined by frontal analysis at 50% breakthrough using copper solution.
⁴ Optimal flow rate during binding is depending on the sample.