VALIDATION IMAGES
Formalin-fixed and paraffin embedded human breast cancer labeled with Anti-CA153/MUC1/KL-6 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.
MCF7 (Positive) and HL-60 (Negative control) cells (black) were fixed with 4% PFA for 10min at room temperature, permeabilized with 20% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with MUC1 Antibody (bs-1497R) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
MCF-7 cells were stained with MUC1 Polyclonal Antibody, Unconjugated(bs-1497R) at 1:100 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with MUC1 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with MUC1 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with MUC1 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with MUC1 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with MUC1 Polyclonal Antibody, Unconjugated (bs-1497R) at 1:200 overnight at 4°C, DAB staining.
MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (MUC1) polyclonal Antibody, Unconjugated (bs-1497R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.