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Sample: Kidney (Mouse) lysate at 30 ug, Primary: Anti-Uromucoid at 1: 300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG at 1: 5000 dilution, Predicted band size: 61/65 kD. Observed band size: 65 kD |
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Sample: Lane1: Kidney (Rat) Lysate at 30 ug. Lane2: Kidney carcinoma (Human) Lysate at 30 ug. Primary: Anti-MCKD2/UMOD at 1: 200 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG at 1: 3000 dilution, Predicted band size: 65kD. Observed band size: 65kD. |
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Tissue/cell: mouse kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-MCKD2/UMOD Polyclonal Antibody, Unconjugated 1: 200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/cell: rat kidney tissue,4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-Uromucoid Polyclonal Antibody, Unconjugated 1: 200, overnight at 4C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1: 200 dilution for 40 minutes at 37C. DAPI (5ug/ml, blue) was used to stain the cell nuclei. |
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Tissue/cell: human kidney tissue,4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-Uromucoid Polyclonal Antibody, Unconjugated 1: 200, overnight at 4C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1: 200 dilution for 40 minutes at 37C. DAPI (5ug/ml, blue) was used to stain the cell nuclei. |
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Blank control: Mouse kidney (blue).Primary Antibody: Rabbit Anti-Coxsackie Adenovirus Receptor antibody (Green), Dilution: 1 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 1 X PBS containing 0.5% BSA. Protocol. The cells were fixed with 2% paraformaldehyde for 10 min at 37C. Primary antibody (1 µg /1x10 6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 10% 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 20, 000 events was performed. |
