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Figure 6 Immunocytochemistry Validation of ASC in HL60 CellsImmunocytochemical analysis of HL60 cells using anti-ASC antibody (2287) at 5 &956,g/ml. Cells 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 5 Immunofluorescence Validation of ASC in Human Spleen TissueImmunofluorescent analysis of 4% paraformaldehyde-fixed Human Spleen Tissue labeling ASC with 2287 at 20 &956,g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). |
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Figure 4 Immunohistochemistry Validation of ASC in Human Spleen Tissue Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ASC antibody (2287) at 2.5 &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 HL60 CellsLoading: 15 &956,g of lysates per lane.Antibodies: ASC 2287, (1 &956,g/mL) in the absence (A) or presence (B) of blocking peptide, 1h incubation at RT 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: ASC 2287, (2 &956,g/mL), ASC 39-001, (2 &956,g/mL), beta-actin (1 &956,g/mL) and GAPDH (0.02 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit or goat anti-mouse (for ASC 39001) IgG HRP conjugate at 1:10000 dilution. |
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Figure 3 Western Blot Validation in Human THP-1 CellsLoading: 15 &956,g of lysate per lane.Antibodies: ASC 2287, (2 &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 ASC in THP-1 Cells (Dowds et al., 2004) Immunofluorescence analysis with anti-ASC antibodies (2287) was performed for BIM in 293 cells transfected with GFP siRNA or ASC siRNA. ASC expression was disrupted after ASC siRNA knockdown. |
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Figure 8 Overexpression Validation of ASC in HEK293T Cells (Dowds et al., 2004) Western blot analysis with anti-ASC antibodies (2287) was performed for ASC in HEK293T cells transfected with pcDNA3-ASC. |