VALIDATION IMAGES
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 CD134 Monoclonal Antibody, Unconjugated (bsm-60298R) 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 CD134 Monoclonal Antibody, Unconjugated (bsm-60298R) 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 Thymus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with CD134 Monoclonal Antibody, Unconjugated (bsm-60298R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
The MJ (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Primary Antibody (green): Rabbit Anti-CD134 antibody (bsm-60298R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed MJ (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (CD134) monoclonal Antibody, unconjugated (bsm-60298R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) 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 CD134 monoclonal antibody, unconjugated (bsm-60298R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.