PureCube Cu-NTA Agarose

Order number: 31403-Cu

€145.00*

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Description

Our PureCube Cu-NTA agarose resins are small agarose resin beads with a diameter of 40 µm. They are used for the purification of active His-tagged proteins from cells and the enrichment of Copper-binding proteins. Our year-long experience in manufacturing agarose resin lead to the high yield of 80 mg protein per ml resin, which is leading in the market compared to other Cu-NTA suppliers. PureCube Cu-NTA resins are suited for batch spin columns and FPLC. Their small diameter provides them with great mechanical stability.

Furthermore, you can also get these Cu-NTA beads pre-packed in a column/cartridge or as magnetic beads.
Feature
Usage Specific binding and purification of 6x His-tagged proteins
Binding of Copper-binding proteins
Specificity Affinity to His-tagged proteins
Affinity to Copper-binding proteins
Binding capacity >80 mg/mL
Bead Ligand Cu-NTA
Bead size 40 μm
Filling quantity Delivered as a 50 % suspension
Metal ion capacity > 15 µeqv Cu2+/ml
pH stability 2-14
Other stabilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
Required equipment
  • Lysis Buffer
  • Wash Buffer
  • Elution Buffer
  • Ice bath
  • Refrigerated centrifuge for 50 mL tube (min 10,000 x g)
  • 50 mL centrifuge tube
  • Micropipettor and Micropipetting tips
  • Disposable gravity flow columns with capped bottom outlet, 2 ml
  • pH meter
  • End-over-end shaker
  • SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment

Lab Results

Different metal ions confer different binding affinity and specificity

Loading different metal ions to a resin results in differing affinity and specificity for a His-tagged protein. Generally, copper exhibits the highest protein yield of commonly used IMAC metal ions, leading to relatively lower purity. In searching for the optimal resin to purify a protein, it is recommended to explore different chelating ligands (IDA or NTA) and different metal ions.
Metal ions compared in their performance for His-tag purifications
Figure 1: Affinity and specificity of metal ions commonly used for IMAC. Loading an IMAC resin with different metal ions can adjust the affinity and specificity to optimize the purity and yield of a purified protein.

Video

Video Guide - How to pack FPLC cartridges


Video Guide - FPLC


Video Guide - Column Chromatography


Video Guide - Batch Spin Chromatography


FAQ

Can I get the datasheet for the Cu-NTA resin?

What are the reasons for non-specific binding?

Some protein can bind to Cu-NTA even without having a His-tag. But to a lesser extent. Washing with NaOH after elution of your protein of interest removes unspecific bound proteins from your resin.

I want to use a high concentration of EDTA and DTT. Is it possible to use Zn-NTA from Cube Biotech?

No, it is not recommended because nickel-ions are reduced with DTT or dissolved with EDTA. If you want to use high concentrations of EDTA and DTT you should use our Indigo resin.

Can Cu-NTA beads be stripped?

Yes, they can. However, we recommend using our pre-stripped NTA or IDA beads, to begin with.

After using DTT my resin changed color. How to regenerate it?

The DTT has probably destroyed your beads. Cu-NTA beads only have a very limited DTT tolerance. However, you can regenerate them to regain their functionality. Please read our detailed protocol for more information regarding this. It is linked above.

However, we would recommend using Ni-INDIGO products instead. They work with the same buffers and protocols as the Cu-NTA products but have a DTT tolerance of 20 mM.