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Blank control: Hepg2 (blue). Primary Antibody: Rabbit Anti-E cadherin antibody (orb501008, Green), dilution: 3 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), dilution: 1:200 in 1X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde for 10 min at 37C. Primary antibody (orb501008, 3 µg/1x10 6 cells) were incubated for 30 min at room temperature, followed by 1X PBS containing 0.5% BSA + 1 0% goat serum (1 hour) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20000 events was performed. |
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Sample: Breast ca (Mouse) Lysate at 40 ug, Primary: Anti-N-cadherin (orb501008) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 100 kD, Observed band size: 105 kD. |
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Sample: Colon carcinoma (Human) lysate at 30 ug, Brain (Rat) lysate at 30 ug, Primary: Anti-CDH2/N-cadherin (orb501008) at 1:200 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:3000 dilution, Predicted band size: 100kD, Observed band size: 100kD. |
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Sample: HepG2 Cell Lysate at 40 ug, Eye (Mouse) Lysate at 40 ug, Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-N-cadherin (orb501008) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 100kD, Observed band size: 110kD. |
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Tissue/Cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-N-cadherin Polyclonal Antibody, Unconjugated (orb501008) 1:400, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: SH-SY5Y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (N-cadherin) polyclonal Antibody, Unconjugated (orb501008) 1:100, 90 minutes at 37C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei. |
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Tissue/Cell: SH-SY5Y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (N-cadherin) polyclonal Antibody, Unconjugated (orb501008) 1:100, 90 minutes at 37C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei. |