PureCube Fe-NTA Agarose

Order number: 31403-Fe

€160.00*

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Description

PureCube Fe-NTA agarose resin beads were developed for the enrichment of phosphorylated proteins. They can also be used for the purification of his tagged proteins as a secondary function. The affinity matrix is based on 7.5% cross-linked agarose that was magnetized. The bead ligand is Fe-NTA and the particle diameter is 40 µm. They are very homogeneous in size, yielding in a high degree of reproducibility.

As an alternative we also offer PureCube Fe-NTA MagBeads and this resin pre-packed in cartridges/columns.

Datasheets

Fe-NTA Agarose Datasheet
Phosphopeptide Enrichment Protocol
This protocol is written for the use of magnetic beads use it as a reference to create a corresponding agarose resin protocol.

Feature
Usage	Enrichment of phosphopeptides Specific binding and purification of 6x his-tagged proteins
Specificity	Affinity to His-tagged proteins Affinity to phosphorylated biomolecules
Bead ligand	Fe-NTA
Bead size	40 µm
Filling quantity	Delivered as a 50 % suspension
Matrix	7.5% crosslinked agarose
Chelator stability	Stable in buffer containing 10 mM DTT and 1 mM EDTA
pH stability	2-14
Other stabilities	100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile

Citations

Year	Author
2023	Zhang R., Hu M., Chen H-N., Wang X., Xia Z., Lui Y., Wang R., Xia X., Shu Y., Du D., Meng W., Qi S., Li Y., Xu H., Zhou Z-G., Dai L.
2023	Gao L., Lu Y., Chen H-N., Li Z., Hu M., Zhang R., Wang X., Xu Z., Gong Y., Wang R., Du D., Hai S., Li S., Su D., Li Y., Xu H., Zhou Z-G., Dai. L.,
2024	Zhang R., Hu M., Liu Y., Li W., Xu Z., He S., Lu Y., Gong Y., Wang X., Hai S., Li S., Qi S., Li Y., Shu Y., Shu Y., Du D., Zhang H., Xu H., Zhou Z., Lei P., Chen H-N., Dai L.,
2024	Tu Y., Yang Q., Tang M., Gao L., Wang Y., Wang J., Liu Z., Li X., Mao L., Jia R.Z., Wang Y., Tang T-S., Xu P., Liu Y., Dai L. Jia D.

Lab Results

Automatic Phosphoproteomics for high-throughput projects

Fe-NTA magnetic beads by Cube Biotech have been proven to increase the speed of a high throughput experiment drastically. Leutert et al. (2019) presented the application of PureCube Fe-NTA MagBeads in a procedure that they named R2-P2, which is short for Rapid-Robotic PhosphoProteomics. They used a KingFisherTM Flex for their robotic runs to fully automize the phosphopeptide enrichment process.

Explanation of an automated phophopeptide enrichment system — **Figure 1:** Schematic depiction of the setup of a R2-P2 assay using a KingFisherTM Flex. The robotic configuration allows for loading of eight different 96-well plates. Each plate can be rotated into position under a 96-pin magnetic head that drops down inside the 96-well plate to release, bind, or agitate the magnetic microspheres in solution. In the first robotic run, peptides are captured from lysates by carboxylated magnetic beads, purified, and eluted by digestion at 37°C. Eluted peptides are dried down and can be resuspended for total proteome analysis by LC-MS/MS and/or for automatic phosphopeptide enrichment. Phosphopeptides are enriched using a second robotic run on the KingFisher^{TM Flex}, using Fe-IMAC, Ti-IMAC, Zr-IMAC, or TiO2 magnetic microspheres, and analyzed by LC-MS/MS to obtain the phosphoproteome.
Source: Leutert et al. (2019)

Superiority over other phosphopeptide enrichment methods

Leutert et al. compared three different types of IMAC beads (including our PureCube Fe-NTA) and TiO2 microspheres. As it can be seen in figure 2 our PureCube Fe-NTA magnetic beads presented themselves to be the best option for phosphopeptide enrichment. With our Fe-NTA MagBeads the most unique phosphopeptides (Fig. 2 A and C) were enriched with the highest efficiency (Figure 2 B).

Performance of different phopshopeptide enrichment matrices and methods compared — **Figure 2:** Comparison of phosphopeptide enrichment performance between four different products/methods. A: Number of unique phosphopeptides identified by the different enrichments (mean +/- SD, n = 3). B: Phosphopeptide enrichment efficiency shown as the fraction of phosphorylated peptides over total peptides (mean +/- SD, n = 3). C: Venn diagram of identified phosphopeptides by the different phosphopeptide enrichment methods.
Source: Leutert et al. (2019)

The best product for the best prize

Analyzing and comparing products from various manufacturers and suppliers can often prove to be a laborious task. The presence of varying concentrations and volumes across different suppliers can lead to confusion when trying to identify the optimal cost-benefit ratio. Recognizing these challenges, Cube Biotech has undertaken the effort to compile a comprehensive overview of prices for the frequently utilized phosphopeptide enrichment products (Fig. 2 C). Our intention is to provide researchers with a clearer understanding of the prevailing pricing landscape in the market, facilitating informed decision-making.

Cube Biotech phopshopeptide Enrichment Beads compared to other competitors — **Figure 3:** Cube Biotech does not only offer the best single product for phosphopeptide enrichment, but also the most prize efficient one. The products of Competitor G are ranging at about half the volume of beads you get for 200 USD in comparison to Cube Biotech. Suspension rates vary only slightly with 25% for the products of Cube Biotech, to 20% for Competitor G.

FAQ

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

Yes. Just click here to download it as a PDF File.

What can I do with Fe-NTA beads?

Fe-NTA beads serve two purposes. First, they can be used as the matrix to bind and thus enrich phophopeptides. Second, they can be used similar to e.g Ni-NTA to purify His-tagged proteins via IMAC.

How does the phosphopeptide enrichment process work?

Use this protocol as a reference. It is written for our PureCube Fe-NTA MagBeads, so you have to make some adjustments to it.

Can I enrich all phosphopeptides using Fe-NTA?

No, this is not possible. Not all phosphopeptides bind to Fe-NTA beads. As shown in figure 2 multiples bead types must be used to cover all phosphopeptides off a cell. Because of that we also offer Ti- Zr- and Al-NTA beads.