25 ug total protein per lane of various lysates (see on figure) probed with TCF4 monoclonal antibody, unconjugated (bsm-62320R) 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 Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) 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 Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) 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 Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) 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 brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Ratbbit IgG antibody (red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with TCF4 Monoclonal Antibody, Unconjugated (bsm-62320R) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Ratbbit IgG antibody (red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.

The U-87MG (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.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-TCF4 antibody (bsm-62320R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.