1 / 1

5. MASS PEAK LIST FROM SPECTRA

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.

nguyet
Download Presentation

5. MASS PEAK LIST FROM SPECTRA

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. 5. MASS PEAK LIST FROM SPECTRA • Automated & Semi-automated Software • Inspect the spectra • CheckMascot-Parameters • Manual PTM-validation • Detect Phosphate-Fingerprints • (Long & Tedious) • For Large-Scale Phospho-analyses Bioinformatics Tools are useful • Variants TiO2 • SIMAC • IMAC • TiO2 1. Chemical Treatments- Sample Conditioning for Phosphoproteomic Analyses • According to the sample amount, characteristics & Phospho-goals • Optimal Phospho-Digestions (Tryp+/- Lys-C; +/-SDS; +/-Urea…) • Avoiding phospho-artefacts • Keeping phosphate-groups 6. In Summary • Combining different Phospho-proteomic • Strategies • Recover the highest number of • phosphorylated peptides/proteins Simple samples,usually No Prefraction Large-scale, Prefraction steps 2. Phosphoenrichments of Mono & Multiply phosphorylated peptides 3. Desalting, cleaning and concentrating phosphopeptides • POROS R2 • (unbonded phospho-) • R3 • &/or Graphite • C18 • R3 -C18 4. Isolating and Identifying Mono &Multiply Phosphorylated Residues Testing different fragmentation modes 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)

More Related