PureCube Co-NTA MagBeads

Order number: 31505

€399.00*

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Description

PureCube Co-NTA magnetic beads / MagBeads were developed by Cube Biotech for His-tag protein purification. Our year-long experience in manufacturing agarose resin lead to the high yield of 30 mg protein per ml resin, which is leading in the market compared to other Co-NTA suppliers.

Co-NTA products by Cube Biotech are also available as agarose resin beads for FPLC and batch spin or as XL magnetic beads for more viscous cell media. For your convenience Cube Biotech also offers its own MagBead separator for use with our 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 30 μm
Filling quantity Delivered as a 25 % suspension
pH stability 2-14
Other stabilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile

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 higest 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 use MagBeads


FAQ

Can I get the datasheet for the Co-NTA MagBeads?

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 Ni-NTA from Cube Biotech?

No, it is not recommended because nickel is reduced with DTT or dissolved with EDTA. If you want to use high concentrations of EDTA and DTT you should use our INDIGO-Ni MagBeads.

What is the capacity at high flow rates for the beads?

If higher flow rates are desired we recommend using beads with bigger diameters. We offer Co-NTA MagBeads with a mean diameter of 30µm and 90µm (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 30µm of 90µm beads, we both have average purification amounts of ~30 µg protein/mL beads. With 400 µm (XL) beads (only agarose resin, not MagBeads), this decreases to ~20 µg/mL.

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 turned orange. How to regenerate it?

The DTT has probably destroyed your beads. Ni-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 Co-NTA products but have a DTT tolerance of 20 mM.