Thermo Scientific™ Pierce™ High pH Reversed-Phase Peptide Fractionation Kit increases protein identification from LC/MS analysis through orthogonal peptide fractionation of complex peptide samples.
• Easy-to-use—resin provided in single-use spin column format
• Improved protein identifications—protein identifications increased by ≥50% when compared to unfractionated samples
• Reproducible—elution profiles and fractional resolution vary by less than 20%
• Optimized—robust procedure for maximal protein identification and peptide recovery while minimizing fractional overlap
• Compatible—reagents validated with a variety of complex samples, including TMT™-labeled peptides
In order to enable deep proteome sequencing, it is often necessary to reduce the sample complexity by fractionation in an orthogonal dimension prior to LC/MS analysis. The Pierce High pH Reversed-Phase Peptide Fractionation Kit utilizes high pH reversed-phase chromatography to separate peptides by hydrophobicity and provides excellent orthogonality to low pH reversed-phase LC-MS gradients.
The kit has been designed to improve protein identification through the use of a proprietary reversed-phase resin in an easy-to-use spin column format with a highpH fractionation protocol. In contrast to strong cation exchange (SCX) fractionation, the high-pH reversed-phase fractions do not require an additional desalting step before LC/MS analysis.
The High pH Reversed-phase Peptide Fractionation Kit includes a high pH buffer (0.1% triethylamine) and twelve spin columns containing pH-resistant reversed-phase resin. Each reversed-phase fractionation spin column enables fractionation of 10–100 µg of peptide sample using a microcentrifuge.
Native, phosphorylated, Tandem Mass TagTM (TMTTM)-labeled, and other complex peptide mixture samples can be fractionated using this kit. Combining the search results generated by the individual fractions helps improve protein sequence coverage and increases the number of identified proteins relative to unfractionated samples.
• Reducing sample complexity to identify targets of interest
• Conducting systems biology studies
• Reducing sample complexity to improve quantitation studies