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Figure 1. ELISA showing titration of anti-NGEP-p1 and p2 antisera (A) and purified anti-NGEP-p1 and p2 antibodies (B) against NGEP-p1 and p2. Western blotting analysis of NGEP protein expression in three prostate tissues (lanes 1, 2 and 3) and a control cell lysate not expressing this protein, DU145 cell (lane 4), detected using affinity purified anti-NGEP-p2 antibody. The expression of NGEP protein was normalized to the expression level of β-actin
(C) . For color figures, please see online at http://www.futuremedicine.com/doi/full/10.2217/BMM.14.106
(Figure omitted. See article PDF.)
Figure 2. Immunohistochemical staining pattern of NGEP expressionin human prostate tissues using anti-NGEP-p2 antibody staining predominantly apical and lateral of epithelial cell membranes. (A) Normal, (B) PCa with Gs; 7(3+4), (C) Pca with Gs; 7(4+3) (original magnification ×200).
(Figure omitted. See article PDF.)
Figure 3. Comparison of NGEP expression in prostate cancer with normalized concentrations of both polyclonal anti-NGEP-p2 antibody (A) and commercialized antibody (B). The prostatectomy of anti-NGEP-p2 antibody on cores was stronger than the commercialized polyclonal antibody to NGEP (original magnification ×100).
(Figure omitted. See article PDF.)
Figure 4. Antibody binding inhibition by preincubation with peptide prior to incubation with prostate tissue. (A) Strong staining of normal tissue by anti-NGEP-p2 antibody (1:20 dilution) was observed in the absence of NGEP-p2. (B) The NGEP-p2 was used (at 2, 5, 10 µg/ml) to block anti-NGEP-p2 antibody (1:20 dilution). No staining was observed, indicating that the anti-NGEP-p2 antibody was specific for the regions of interest.
(Figure omitted. See article PDF.)
Figure 5. Immunohistochemical analysis of NGEP expression in cores (A & B), needle biopsy (C) and lymph node tissue (D) using anti-NGEP-p2 antibody. (A & B) Strong intensity of NGEP in PCa with low grade tumor and BPH, respectively. (C) Tumor tissue of needle biopsy specimen with high grade stained weak and (D) Lymph node tissue associated with tumor metastasis stained weak (original magnification ×100 [A-C] and ×200 [F] ). (E) The average of the intensity of NGEP expression in normal prostate tissues and PCa including low grade and high grade (Mann-Whitney U test).
(Figure omitted. See article PDF.)
Figure 6. Staining several normal tissues using anti-NGEP-p2 antibody. No cross-reactivity was observed in normal tissues of the brain (A) , heart (B) , lung (C) , liver (D) and kidney (E)