PureCube Co-NTA Agarose

Order number: 31403

€146.00*

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Description

Our PureCube Co-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. Our year-long experience in manufacturing agarose resin lead to the high yield of 40 mg protein per ml resin, which is leading in the market compared to other Co-NTA suppliers. PureCube Co-NTA resins are suited for batch spin columns and FPLC. Their small diameter provides them with great mechanical stability.

For higher flow rates we recommend our Co-NTA agarose beads with 100 µm diameter and for extreme cases our XL-sized Co-NTA beads.
Furthermore, you can also get these beads pre-packed in a column/cartridge or as magnetic beads.
Feature
Usage Specific binding and purification of 6x His tagged proteins
Specificity Affinity to His tagged proteins
Binding capacity >30 mg/mL
Bead Ligand Co-NTA
Bead size 40 μm
Filling quantity Delivered as a 50 % suspension
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, cobalt exhibits the highest binding specificity of commonly used IMAC metal ions, leading to relatively low yields but high purity. Copper, at the other end of the spectrum, has a high affinity leading to high yields but unspecific binding. 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 Co-NTA resin?

Why should I use Co-NTA beads instead of Ni-NTA?

As figure 1 in the "lab results" section illustrates, Co-NTA purifications using the His-tag are purer than Ni-NTA in comparison. This comes at a cost of some of the protein yield. If your needs fit that pattern use Co-NTA beads instead of Ni-NTA.

What are the reasons for non-specific binding?

Even though Co-NTA is the purest form of His-tag purification, some unwanted proteins may still bind to the beads. But 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 Co-NTA from Cube Biotech?

No, it is not recommended because cobalt-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.

How is the capacity at high flow rates?

If higher flow rates are desired we recommend using beads with bigger diameters. We offer Co-NTA beads with mean diameters of 40µm, 100µm, and 400µm (XL).

With each size increase, the flow rates also increase due to the proportionally increasing space between the beads. However, the surface of the beads does not increase at the same speed as the diameter (square-cube-law). That results in decreasing amounts of purified protein per mL beads while increasing the bead sizes.

For 40µm of 100µm beads, we both have average purification amounts of ~30 µg protein/mL beads. With 400 µm (XL) beads, this decreases to an unspecified amount.

We recommend reading the corresponding section of the "Introduction to agarose matrixes" guide on this subject for more detailed information.

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

The DTT has probably destroyed your beads. Co-NTA beads only have a limited DTT tolerance of about 1 mM. However, you can regenerate them to regain their functionality. Please read our detailed protocol for more information regarding this.

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