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PureCube 100 NTA Agarose
PureCube NTA (nitrilotriacetic acid) agarose resin was developed for IMAC based protein purification and protein enrichment methods. Since the affinity of an NTA-metal-ion-complex depends on the loaded metal ion, we offer NTA agarose resin 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 resin is also available with a bead diameter of 40 µm or for even higher FPLC flow rates or XL sized . Alternatively we also offer NTA magnetic beads. Furthermore PureCube NTA agarose (40 µm) can also be obtained pre-filled into FPLC cartridges / columns.
1. Protein Binding Capacity
The protein binding capacity is up to 80 mg/mL, as determined by purification of 6xHis-tagged GFP protein from E.coli cleared lysates, and quantified via spectrophotometry. Proteins are eluted with high purity.
2. Compatibility
PureCube 100 NTA Agarose is very stable and can resist the following conditions in most situations: pH 3-13, 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile, 10 mM DTT, 1 mM EDTA, 1 M NaOH.
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: | Agarose |
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 | 100 μm(Agarose) |
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 |
Other stabilities | 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile |
Filling quantity | Delivered as a 50 % suspension |
Order number Products Bead size Protein Binding Capacity Shipment Temperature Storage temperature Usage Price(yuan) 74703 PureCube 100 NTA Agarose(10ml) 100 Depending on the binding metal Ambient temperature 4° C Excessive metal ions can be selected for the purification of His labeled recombinant proteins; Stable in buffer containing 10 mM DTT, 1 mM EDTA and 1 M NaOH; Maximum pressure: up to 300 kPa; pH Stability: 3-13; Max. Flow Rate: 1000cm/h 1165 74705 PureCube 100 NTA Agarose(50ml) 100 Ambient temperature 4° C 4701 74710 PureCube 100 NTA Agarose(250ml) 100 Ambient temperature 4° C 20290 74712 PureCube 100 NTA Agarose(500ml) 100 Ambient temperature 4° C 36317
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