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PureCube NTA Magbeads XL
PureCube NTA MagBeads / magnetic beads XL were developed for IMAC based purification and enrichment methods. Since the affinity of an NTA-metal-ion-complex depends on the loaded metal ion, we offer NTA magnetic beads uncoupled to any metal ion for the case that we do not offer your desired NTA + metal ion combination. By providing our coupling protocols, feel free to create any NTA purification resin you desire. PureCube NTA MagBeads are also available regular sized (30 µm) for higher protein purifications of more viscous cell lysates or as NTA agarose resin. Furthermore PureCube NTA agarose can also be obtained pre-filled into FPLC cartridges / columns.
1. Protein Binding Capacity
The protein binding capacity is ~40 mg protein per mL of settled beads, as determined by purification of 6xHis-tagged GFP protein from E.coli cleared lysates, and quantified via spectrophotometry.
2. Compatibility
PureCube NTA MagBeads XL are very stable and can resist the following conditions in most situations: pH 2-4, 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile.
3. Different metal ions confer different binding affinity and specificity
His-tag purification is based on ionic interactions of a coupled metal ion and the Poly-His-tag. Therefore other metal ions can also fullfill the function of Nickel. Figure 1 shows the most frequently used metal ions for His-tag purification. Depending on the used metal ion the balance between the specificity and the affinity of the metal ion to the His-tag shifts. Nickel is a balanced choice between both extremes. Copper has the highest affinity, but Cobalt is the choice for the most pure His-tag purifications.
Fig. 1: Depending on which metal ion is coupled to NTA the balance between protein yield and purification specificity shifts. Nickel is a very balanced choice between the two extremes.
Type | Magnetic Beads |
Ligand | NTA |
Coupled Ion | none |
Usage | For coupling of metal ions for corresponding purification and / or enrichment purposes |
Specifity | Varies |
Binding capacity | Depends on the loaded ion |
Purity | Depends on the loaded ion |
Bead Ligand | NTA |
Bead size | ~90 μm(XL MagBeads) |
Matrix | 6% cross-linked agarose |
Recommended FPLC flow rates | 0.5-2 mL/min |
Maximal FPLC flow rate | 6 mL/min |
pH stability | 2-14 |