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. 2015 Oct 20;6(41):43743–43758. doi: 10.18632/oncotarget.6193

Figure 7. Quantitation of extracellular CDCP1 between high- and low-risk prostate cancer.

Figure 7

(A) Detection of CDCP1 in urine-EPS samples. Western blot analysis of urine-EPS samples from low-risk, intermediate-risk, and high-risk PCa were performed with a goat antibody against the extracellular domain (aa 33–666) of CDCP1 (AF2666). (B) Scheme for immune-enriched quantitative mass spectrometry analysis of CDCP1 proteins from urine-EPS. CDCP1 was immunoprecipitated with anti-CDCP1 (AF2666) from urine-EPS as described. Goat IgG was included as an isotype control. Eluents of the purified CDCP1 proteins were subjected to SDS-PAGE. Gel bands were cut into 15 equally spaced pieces (25–150 kDa) and subjected to LC-MS/MS analysis. (C) Identification of peptides spanning extracellular domains of CDCP1 detected by LC-MS/MS. (D) Representative MS/MS spectrum of tryptic peptide TFIWDVK of sCDCP1 digestion. (E) Quantitative precursor MS results display simultaneous identification and quantitation results for the 6 unique peptides from CDCP1. The left panel shows the relative peak area differences for the combined targeted peptides. The right panel shows the extracted ion chromatogram and integrated peak area of primary ions per targeted peptide.