PureCube Ti-NTA MagBeads

Order number: 31501-Ti

€94.00*

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Description

PureCube Ti-NTA Magbeads / Magnetic 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 6% cross-linked agarose that was magnetized. The bead ligand is Ti-NTA and the particle diameter is 30 µm. They are very homogeneous in size, yielding in a high degree of reproducibility. The phosphopeptide enrichment with Cube Biotech MagBeads can be automated as it has already been proven in this paper.

Datasheets

Feature
Usage	Enrichment of phosphopeptides
Specificity	Affinity to phosphorylated biomolecules
Bead ligand	Ti-NTA
Bead size	30 µm
Filling quantity	Delivered as a 25 % suspension
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
2024	Schmidt L., Saynisch M., Hoegsbjerg C., Schmidt A., MacKey A., Lackmann J-W., Müller S., Koch M., Brachvogel B., Kjaer M., Antczak P., Krüger M.

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 (see 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 Ti-NTA MagBeads?

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

What can I do with Zr-NTA beads?

They are meant to be used as the matrix to bind and thus enrich phophopeptides.

How does the phosphopeptide enrichment process work?

It is quite simple. Just follow this protocol.

Can I enrich all phosphopeptides using Ti-NTA?

No, this is not possible. Not all phosphopeptides bind to Ti-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 Fe-, Zr- and Al-NTA beads.