Uromucoid Polyclonal Antibody

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

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.

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 37°C for 20 min; Incubation: Anti-MCKD2/UMOD Polyclonal Antibody, Unconjugated 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

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 37°C for 20 min; Incubation: Anti-Uromucoid Polyclonal Antibody, Unconjugated 1: 200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1: 200 dilution for 40 minutes at 37°C. DAPI (5ug/ml, blue) was used to stain the cell nuclei.

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 37°C for 20 min; Incubation: Anti-Uromucoid Polyclonal Antibody, Unconjugated 1: 200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1: 200 dilution for 40 minutes at 37°C. DAPI (5ug/ml, blue) was used to stain the cell nuclei.

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 37°C. 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.