Application
Fig1: Tissue/cell: mouse kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-MCKD2/UMOD Polyclonal Antibody, Unconjugated(ER1918-44) 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Application
Fig2: Sample:
Lane 1: Kidney(Rat) Lysate at 30 ug
Lane 2: Kidney carcinoma(Human) Lysate at 30 ug
Primary: Anti-MCKD2/UMOD (ER1918-44) at 1:200 dilution;
Secondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 3000 dilution;
Predicted band size : 65kD
Observed band size : 65kD
Application
Fig3: Sample: Kidney(Mouse) lysate at 30 ug;
Primary: Anti-Uromucoid (ER1918-44) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-rabbit IgG(bs-0295G-HRP) at 1: 5000 dilution;
Predicted band size:61/65 kD
Observed band size:65 kD
Application
Fig4: Blank control: Mouse kidney (blue).
Primary Antibody:Rabbit Anti-Coxsackie Adenovirus Receptor antibody (ER1918-44,Green); Dilution: 1μg 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℃. Primary antibody (ER1918-44, 1μg /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.