TiO2 magnetic particles for high throughput phosphopeptide isolation.

The Thermo Scientific Pierce Magnetic Titanium Dioxide Phosphopeptide Enrichment Kit is for isolating phosphopeptides from complex biological samples using titanium dioxide-coated magnetic beads. The TiO2 ligand selectively binds peptides containing phosphorylated serine (Ser), tyrosine (Tyr) or threonine (Thr), enabling phosphopeptide enrichment from protease-digested samples. The isolated phosphopeptides are compatible for analysis downstream by mass spectrometry.

The high-performance, iron oxide, superparamagnetic particles are validated and optimized for use with high-throughput magnetic platforms, such as the Thermo Scientific KingFisher 96 and KingFisher Flex Instruments. The beads also enable premium performance for simple benchtop applications using an appropriate magnetic stand.

Learn more about automated magnetic separations and Thermo Scientific Automated Solutions.

Highlights:

Complete MS-compatible Kits – include ready-to-use binding, wash and elution buffers that are optimized for phosphopeptide enrichment and downstream analysis by MALDI and ESI mass spectrometry
Optimized for HTS – procedure validated for processing 1 to 96 samples at a time; complete entire assay in about 15 minutes using a KingFisher Flex Instrument
Stable affinity ligand – titanium dioxide is specially coated as a film on the magnetic particles
Selective – affinity system is selective for phosphorylated Ser, Tyr and Thr; exhibits minimal non-specific binding to acidic residues
Sensitive – affinity provides more than 1000 times greater sensitivity than traditional IMAC technologies; enables enrichment and MS-measurement of less than 100 fmol of phosphoprotein

Phosphopeptide enrichment improves MS-identification of phosphoproteins. Two milligrams of a tryptic digest prepared from periferal blood mononuclear cells (lymphocytes) with and without phosphopeptide enrichment were analyzed by MS. Enrichment was performed with the Thermo Scientific Pierce Titanium Dioxide Phosphopeptide Enrichment Kit using the Thermo Scientific KingFisher 96 Instrument. Samples were analyzed on an Thermo Scientific LTQ Orbitrap Mass Spectrometer.
	Enriched	Non-Enriched
Total number of proteins identified	185	247
Total number of phosphoproteins identified	160	1
Total number of peptides identified	2347	2457
Total number of phosphopeptides identified	2009	7
Total number of unique phosphopeptides identified	177	1
Relative enrichment for phosphopeptides (%)	86	0.3

Product #	Description	Pkg. Size
88811	Pierce Magnetic Titanium Dioxide Phosphopeptide Enrichment Kit Includes: TiO₂ Magnetic Beads (20X), 1 ml Binding Buffer, 100 ml Washing Buffer, 25 ml Elution Buffer, 3 ml Thermo-Fast 96 Robotic PCR Plate (0.2 ml wells), 2 plates	Kit
88812	Pierce Magnetic Titanium Dioxide Phosphopeptide Enrichment Kit, Trial Size Includes: TiO₂ Magnetic Beads (20X), 0.25 ml Binding Buffer, 100 ml Washing Buffer, 25 ml Elution Buffer, 3 ml Thermo-Fast 96 Robotic PCR Plate (0.2 ml wells), 2 plates	Kit