VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded Human Skin tissue; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 15 minutes; Blocking buffer (3% BSA) at room temperature for 30min; Antibody incubation with CD44 (Hermes-3) Monoclonal Antibody (bsm-51065M) at 1:25 for 1 hour at 37°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human lung adenocarcinoma section; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 15 minutes; Blocking buffer (3% BSA) at room temperature for 30min; Antibody incubation with CD44 (Hermes-3) Monoclonal Antibody (bsm-51065M) at 1:25 for 1 hour at 37°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded SAMPLE tissue; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 15 minutes; Blocking buffer (3% BSA) at room temperature for 30min; Antibody incubation with CD44 (Hermes-3) Monoclonal Antibody (bsm-51065M) at 1:50 for 1 hour at 37°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.
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 CD44 (H3) Monoclonal Antibody, Unconjugated (bsm-51065M) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human tonsil; 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 CD44 (H3) Monoclonal Antibody, Unconjugated (bsm-51065M) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human pancreatic 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 CD44 (H3) Monoclonal Antibody, Unconjugated (bsm-51065M) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Human MDA-MB-231 cell lysates; Lane 2: Human U251 cell lysates; Lane 3: Human A549 cell lysates probed with CD44 Monoclonal Antibody, Unconjugated (bsm-51065M ) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Hela cells were incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with CD44 (H3) Monoclonal Antibody(bsm-51065M)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
25 ug total protein per lane of various lysates (see on figure) probed with CD44 monoclonal antibody, unconjugated (bsm-51065M) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.