Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Phosphopeptide Enrichment Using Offline Titanium Dioxide Columns for Phosphoproteomics

doi: 10.1007/978-1-62703-360-2_8

Figure Lengend Snippet: Nanoflow reversed-phase liquid chromatography coupled online with a linear ion trap mass spectrometer for the analysis of TiO2-enriched phosphopeptides from a HeLa cell lysate using collision-induced dissociation (CID) for tandem MS and phosphate neutral loss-dependent MS/MS/MS (RPLC-MS2-MS3). The analysis is for 5 μL injection of phosphopeptides onto a 75 μm ID capillary column. The lower panel is the base peak chromatogram of the analysis. The upper panel shows the MS3 or MS2 spectra of the phosphopeptides ALVAT*PGKK, SGAQASSTPLS*PTR, and LLT*PTHSFLAR identified, respectively, from nucleolin, lamin-A/C, and microtubule-associated protein 7. The relatively high abundance levels of the LC chromatographic peaks, from which these phosphopeptides were eluted, demonstrate that phosphopeptides were effectively enriched from the HeLa cell lysate. The asterisk marks the phosphorylated Ser or Thr residues.

Article Snippet: Cell Culture of HeLa Cells Human cervical epithelial cancer cell line HeLa (ATCC).

Techniques: Reversed-phase Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Injection

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Phosphopeptide Enrichment Using Offline Titanium Dioxide Columns for Phosphoproteomics

doi: 10.1007/978-1-62703-360-2_8

Figure Lengend Snippet: Nanoflow reversed-phase liquid chromatography coupled online with a linear ion trap mass spectrometer for the analysis of TiO2-enriched phosphopeptides from a HeLa cell lysate using collision-induced dissociation (CID) for tandem MS and phosphate neutral loss-dependent MS/MS/MS (RPLC-MS2-MS3). The analysis is for 5 μL injection of phosphopeptides onto a 75 μm ID capillary column. The lower panel is the base peak chromatogram of the analysis. The upper panel shows the MS3 or MS2 spectra of the phosphopeptides ALVAT*PGKK, SGAQASSTPLS*PTR, and LLT*PTHSFLAR identified, respectively, from nucleolin, lamin-A/C, and microtubule-associated protein 7. The relatively high abundance levels of the LC chromatographic peaks, from which these phosphopeptides were eluted, demonstrate that phosphopeptides were effectively enriched from the HeLa cell lysate. The asterisk marks the phosphorylated Ser or Thr residues.

Article Snippet: Human cervical epithelial cancer cell line HeLa (ATCC).

Techniques: Reversed-phase Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Injection