VALIDATION IMAGES
The HepG2 (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-FSCN1 antibody (bsm-61626R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed HepG2 (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (FSCN1) monoclonal Antibody, unconjugated (bsm-61626R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
25 ug total protein per lane of various lysates (see on figure) probed with FSCN1 monoclonal antibody, unconjugated (bsm-61626R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with FSCN1 Monoclonal Antibody, Unconjugated (bsm-61626R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.