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Figure 4 Immunofluorescence Validation of TIM-1 in Human Uterus CellsImmunofluorescent analysis of 4% paraformaldehyde-fixed human uterus cells labeling TIM-1 with 3809 at 20 &956,g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing both membrane and cytoplasmic staining on human uterus cells. |
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Figure 5 Immunohistochemistry Validation of TIM-1 in Human Uterus TissueImmunohistochemical analysis of paraffin-embedded human uterus tissue using anti-TIM-1 antibody (3809) at 10 &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 1 Western Blot Validation in Human Cell LinesLoading: 15 &956,g of lysates per lane.Antibodies: TIM-1 3809 (8 &956,g/mL), overnight incubation at 4&730, C in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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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: TIM-1 3809 (8 &956,g/mL), TIM-1 3811 (1 &956,g/mL), beta-actin (1 &956,g/mL), and GAPDH (0.02 &956,g/mL), overnight incubation at 4&730, C (3809) or 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 3 Validation with TIM-1 siRNA Knockdown in Hela CellsHeLa cells were transfected with control siRNAs (lane 1) or TIM-1 siRNAs (lane 2) Loading: 15 &956,g of HeLa whole cell lysates per lane.Antibodies: 3809 (8 &956,g/mL), 1 h 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 the Mouse Cell LineLoading: 15 &956,g of NIH/3T3 cell lysates.Antibodies: TIM-1 3809 (8 &956,g/mL), overnight incubation at 4&730, C in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |