PureCube Ni-NTA Agarose

The His tag is the most widely used affinity tag due to its small size, low immunogenicity, and versatility under native or denaturing conditions, as well as in presence of detergents and many other additives.

 

Cube Biotech offers high-performance PureCube Ni-NTA Agarose with 40 µm average bead size, based on BioWorks Workbeads, for purification of his-tagged proteins. 

 

PureCube Ni-NTA Agarose is provided as a 50% suspension, and also available as prepacked chromatography columns. For purification of his-tagged proteins from cell culture supernatants or for pull-down experiments, we recommend PureCube Ni-NTA MagBeads.

 

To detect His-tagged proteins in Western Blot experiments, Cube Biotech offers the highly specific PentaHis antibody.

 

Do you prefer Ni-IDA? We offer the same resin with that chelating ligand! (what's the difference?)

 

PureCube Affinity Resins

PureCube His Affinity ResinsPureCube Ni-NTA Agarose

 

 

Why PureCube Ni-NTA Agarose?

  High binding capacity (up to 80 mg/mL) and high purity

  Stable in 10 mM DTT and 1 mM EDTA. Need to use higher concentrations?

  40 µm agarose beads. Need larger beads for higher flow rates?

  Also available loaded with Co, Cu, Zn, and other metals

 

-> Unsure which His affinity product to choose? See our overview.

 

 

 

Ni-NTA Agarose was successfully used in the following publications:

 

Hsieh, Y.-L. et al.: Molecular Characterization of Ethylene Response Sensor 1 (BoERS1) in Bambusa oldhamii.

Plant Mol Biol Rep, Published online August 21, 2015

 

Stressler, T. et al.: A Novel Glutamyl (Aspartyl-) Specific Aminopeptidase A from Lactobacillus delbrueckii with Promising Properties for Application.

PLOS ONE, DOI:10:1371/journal.pone.0152139 March 22, 2016.

 

Hai-Chao Li  et al.: A New Homo-Hexamer Mn-Containing Catalase from Geobacillus sp. WCH70. 

Catalysts 2017, 7(9), 277; doi:10.3390/catal7090277, September 18, 2017

 

 

 

Over 20% more yield obtained with PureCube Ni-NTA Agarose

Fig. 1: Over 20% more yield obtained with PureCube Ni-NTA Agarose. SDS-PAGE of GFP expressed in E. coli and purified in gravity columns with PureCube Ni-NTA Agarose and Ni-NTA resin from Competitor Q. 80 mg/mL protein yield was obtained with PureCube Ni-NTA Agarose (E1–E4, Cube) compared to 65 and 48 mg/mL, respectively, with the widely used alternative resins G and Q (E1–E4, Competitor G / Competitor Q).

 

High yield and purity

Our unique production process yields a Ni-NTA Agarose that exhibits a protein binding capacity >20% higher than that of two leading competitor products. Figure 1 shows the SDS-PAGE of GFP expressed in E. coli and purified in gravity colums with PureCube Ni-NTA Agarose and the Ni-NTA resin from Competitor G and Competitor Q. The protein yield in 4 elutions (E1-E4, Cube) was 80 mg/mL, compared to 65 and 48 mg/mL obtained with the alternative resins (E1-E4, Competitor G, Competitor Q). Similar results (10-18% higher binding capacity; data not shown here) were obtained comparing the purification of JNK1 (Kinase, 48 kDa) on PureCube Ni-NTA and the Ni-NTA of leading providers.

 

 

NTA is robust in the presence of reducing and chelating agents (DTT / EDTA)

Fig. 2: NTA is robust in the presence of reducing and chelating agents. GFP-His was purified on gravity columns containing PureCube Ni-NTA Agarose after exposing the resin for 1 h to 3 concentrations of DTT or EDTA. NTA exhibits a shallow decay rate in binding capacity.

 

 

Superior DTT and EDTA stability

PureCube Ni-NTA Agarose is very robust in the presence of DTT and EDTA. In a stability test, PureCube Ni-NTA Agarose was exposed to increasing concentrations of DTT or EDTA for

1 h. Thereafter, the resins were used to purify E. coli-expressed GFP-His in gravity columns. The binding capacity of the resin decreased in the presence of both DTT and EDTA but the decay rate was shallow. In presence of DTT, PureCube Ni-NTA Agarose lost on average 8% binding capacity with each increase in DTT concentration, resulting in an overall decay of 22% at 10 mM. Even at 1.5 mM EDTA, the resin still exihibits 54% of its maximum binding capacity (Fig. 2).

 

 

PureCube Ni-NTA Agarose is robust against oxidation and regenerable.

Fig. 3: PureCube Ni-NTA Agarose is robust against oxidation and regenerable. PureCube Ni-NTA Agarose was exposed to 5mM DTT for 1 h (A). After demonstrating that it could still bind GFP (B), the resin was washed, stripped (C), and reloaded with Ni2+ (D) following standard Cube protocol (see Cube Protocols & Datasheets).

 

Robust against oxidation and regenerable

PureCube Ni-NTA Agarose retains its color and function after exposure to as much as 10 mM DTT. Figure 3 shows a photo series of the resin after a 1 h exposure to 5 mM DTT. Unlike other resins, PureCube Ni-NTA Agarose did not turn brown (A). The resin was still able to bind GFP (B), with a measured binding capacity of 65 mg/mL (see Fig. 2). The resin could then be regenerated by stripping the NTA, turning the resin white (C), and reloading it with nickel ions (D). The protocol for regenerating PureCube Ni-NTA Agarose can be downloaded.

 

 

  From Oct 1-Dec 31, 2017, buy this product with a 20% discount.

Quote "Cube2017" when placing your order. Learn more

 

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Ni-NTA Agarose

PureCube Ni-NTA Agarose
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PureCube Ni-NTA Agarose (10 ml)

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PureCube Ni-NTA Agarose (50 ml)

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PureCube Ni-NTA Agarose (250 ml)

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