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Titansphere
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Protein phosphorylation is a fundamental process and influences cell differentiation, growth and migration. However, analysing phosphorylated proteins is very complicated due to the low concentrations and low ionisation efficiency in MS analyses. It is therefore important to concentrate the phosphorylated proteins and reduce the non-phosphorylated proteins in order to improve the analysis of protein digest studies using LC/MS.
The Titansphere products from GL-Sciences have been specially developed for this application. Titansphere products are more efficient than IMAC (Immobilised metal affinity chromatography) purification, whereby both methods can be combined for optimal results. The phosphate group of a protein is absorbed on the surface of the Titansphere particles under acidic conditions and eluted under basic conditions. Non-phosphorylated proteins do not absorb and are therefore not retained. This means that purification can take place under acidic conditions. The Titansphere material is available in bulk for various applications. The advanced form Titansphere Phos-TiO is available in bulk as well as Spin Tips and Titansphere Phos-TiO MP Kit.
Product Line
- Inertsil
- InertSustain
- InertSustain Swift
- InertCap
- InertSep
- Titansphere
- LD239 Gas Leak Detector
- GF-1010 Digital Gas Flow Meter
- Bioptic
- GL Sciences-HayeSep
- GL Sciences-Porapak
- GL Sciences-TC
- GL Sciences-Tenax
- GL Sciences-Unibeads
- InertCore
- InertSep WAX FF/GCB
- InertSphere
- MonoCap
- MonoFas
- MonoSpin
- MonoTrap
- OPTIC
- ProteoSil
- SYPRON
Technical Data
Titansphere bulk material
| Description | Titansphere TiO |
| Particle size | 5, 10 µm |
| Particle shape | Spherical |
| Absorption point | Titanium dioxide crystals |
| Pore size | 100 Å |
| pH range | 2.0 - 12.0 |
| Description | Particle size | Quantity | Art. No. |
| Titansphere TiO | 5 µm | 500 mg | 5020-75000 |
| Titansphere TiO | 10 µm | 500 mg | 5020-75010 |
Titansphere Phos-TiO bulk material
| Description | Particle size | Quantity | Art. No. |
| Titansphere Phos-TiO Bulk | 10 µm | 500 mg | 5010-21315 |
Titansphere Phos-TiO Kit
| Description | 5010-21309 | 5010-21310 | 5010-21311 | 5010-21312 |
| Titansphere Phos-Tio Kit | 24 pcs | 96 pcs | 24 pcs | 96 pcs |
| Titansphere Sorbent Mass/Tip Volume | 1mg/10µl | 1mg/10µl | 3mg/200µl | 3mg/200µl |
| Spin Tip Quantity | 24 pcs | 96 pcs | 24 pcs | 96 pcs |
| Waste Tube quantity | 24 pcs | 96 pcs | 24 pcs | 96 pcs |
| Recovery tube quantity | 24 pcs | 96 pcs | 24 pcs | 96 pcs |
| Instructions | 1 pc | 1 pc | 1 pc | 1 pc |
Titansphere Phos-TiO MP Kit
| Description | 5010-21282 | 5010-21283 |
| Titansphere Phos-TiO MP Kit | 24 pcs | 24 pcs |
| Titansphere Sorbent Mass/Tip Volume | 1 mg / 200 μL | 3 mg / 200 μL |
| Spin Tip Quantity | 24 pcs (6 x 4 packs) | 24 pcs (6 x 4 packs) |
| Waste Fluid Tube Quantity | 24 pcs | 24 pcs |
| Recovery Tube (2.0 mL) Quantity | 24 pcs | 48 pcs |
| Recovery Tube (1.5 mL) Quantity | 48 pcs | 48 pcs |
| Solution B (Lactic acid) Quantity | 2 mL | 2 mL xb14 |
| Instruction Manual | 1/pk | 1/pk |
Titansphere Phos-TiO MP Kit Protocol
Applications
Efficient purification of HeLa Cell lysate with Titansphere TiO
The HeLa Cell Lysate consists mainly of non-phosphorylated peptides. Titansphere TiO shows excellent selectivity. 90% of the bound peptides are phosphorylated.
Probe: HeLa Cell Lysat, 5µg
Titansphere TiO: 1mg
Comparison of Titansphere TiO with IMAC
Compared to the IMAC method, Titansphere TiO shows a much higher capacity and a significantly higher number of isolated phosphopeptides.
Probe: Arabidopsis Cell Extrakt, 100µg
Titansphere TiO: 1mg
Comparison of Titansphere Tio Bulk and Titansphere Phos-TiO Kit
The Titansphere Phos-Tio kit uses the optimised Titansphere Phos-TiO bulk material. The Phos-TiO material was developed with a view to better absorption and capacity of phosphopeptides. The Titansphere Phos-TiO kit shows a 2.6 times higher peak area and 1.6 times more identified phosphopeptides.
Probe: HeLa Cell Lysat, 50µm
Titansphere Media: 1mg
| Produkt | Phosphoryliert | Nicht Phosphoryliert |
| Titansphere Phos-TiO Kit | 996 | 185 |
| Titansphere TiO Bulk | 635 | 71 |
New TiO2-HAMMOC method vs. SIMAC method with the Titansphere Phos-TiO MP Kit
The working group led by Wakabayashi et al. (2015) has demonstrated the highly efficient recovery of single and multiply phosphorylated peptides using HeLa cells 100µg in contrast to the Sequential Elution from IMAC (SIMAC) method.
By using basic elution buffers, multiply phosphorylated peptides can be recovered.
Wakabayashi M, Kyono Y, Sugiyama N, Ishihama Y (2015): Extended coverage of singly and multiply phosphorylated peptides from a single titanium dioxide microcolumn. Analytical Chemistry 87 (20), 10213-10221.
Conclusion
The new Titansphere Phos-TiO MP Kit enables a higher recovery of single and multiply phosphorylated peptides. This can reduce ion suppression in subsequent LC-MS/MS detection. This kit shows higher capacities for phosphorylated proteins and peptides than the previous Titansphere Phos-TiO kit.
Downloads
Brochures
Gl Sciences Titansphere Phos-TiO MP Kit Brochure
GL Sciences Bioanalysis Catalogue
GL Sciences Phosphepeptide Purifying & Enrichment Products Brochure
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