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Figure 1 Western Blot Validation in (A) Human Raji Cells , (B) Mouse testis tissue lysate and (C) Rat testis tissue lysateLoading: 15 &956,g of lysates per lane.Antibodies: PHAP I 3151 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 5 Immunofluorescence Validation of PHAP I in Mouse Small Intestine cellsImmunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Small Intestine Cells labeling PHAP I with 3151 at 20 &956,g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). |
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Figure 6 Immunohistochemistry Validation of PHAP I in Mouse Small Intestine Tissue Immunohistochemical analysis of paraffin-embedded Mouse Small Intestine Tissue using anti-PHAP I antibody (3151) at 2 &956,g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT, antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4&730,C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
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Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell LinesLoading: 15 &956,g of lysates per lane.Antibodies: PHAP I 3145 (2 &956,g/mL), PHAP I 3151 (1 &956,g/mL), and beta-actin (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 7 KD Validation of PHAPI in Human Breast Cancer Cells (Schafer et al., 2006) Human breast cancer cells (T47D cells) were transfected with control or PHAPI siRNA duplex. PHAPI was detected via Western Blot analysis by using the anti-PHAPI antibody. PHAPI expression was reduced after PHAPI siRNA knockdown. |
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Figure 8 Increased Expression Validation of PHAPI in Patient Samples of BreastTumor Tissue (Schafer et al., 2006) PHAPI was overexpressed in all breast tumor samples of patients and human breast cancer cells (MDA-MB-453), but not in the normal breast tissue or human primary mammary epithelial cells (HMEC). |
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Figure 9 Overexpression of PHAPI in Breast Cancer Cells (Schafer et al., 2006) Western blot analysis with anti-PHAPI antibodies was performed for PHAPI in human cell lines from breast, prostate and lung. PHAPI was overexpressed in breast cancer cells when compared with normal cells (HMEC) whereas there were no significant differences in PHAPI expression in normal and cancer cells of either prostate or lung origin. |
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Figure 3 Western Blot Validation in Human, Mouse and Rat Cell LinesLoading: 15 &956,g of lysates per lane.Antibodies: PHAP I 3151 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 4 Western Blot Validation in Mouse TissuesLoading: 15 &956,g of lysates per lane.Antibodies: PHAP I 3151 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |