5. MASS PEAK LIST FROM SPECTRA. Automated & Semi-automated Software Inspect the spectra Check Mascot-Parameters. Manual PTM-validation Detect Phosphate-Fingerprints (Long & Tedious) For Large-Scale Phospho-analyses Bioinformatics Tools are useful. Variants TiO2. SIMAC. IMAC.
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1. Chemical Treatments- Sample Conditioning for Phosphoproteomic Analyses
6. In Summary
Simple samples,usually No Prefraction
Large-scale, Prefraction steps
2. Phosphoenrichments of Mono & Multiply phosphorylated peptides
3. Desalting, cleaning and concentrating phosphopeptides
4. Isolating and Identifying Mono &Multiply Phosphorylated Residues
MS2, MS3, MS3 NL, MSA
(eg. Combining different ionizations via LC-ESI: CID, ECD,ETD)
Testing different Ph-elutions
DHB, Glycolic & Phtalic acids…
(eg. Check via MALDI-MS, MS/MS)