VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded (Human ovarian 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; Incubation with (Progesterone Receptor) Monoclonal Antibody, Unconjugated (bsm-60804R) at 1:300 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation Progesterone Receptor Monoclonal Antibody, Unconjugated(bsm-60804R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
25 ug total protein per lane of various lysates (see on figure) probed with Progesterone Receptor monoclonal antibody, unconjugated (bsm-60804R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
The T47-D (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-Progesterone Receptor antibody (bsm-60804R): 1 μg/10^6 cells; Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed T-47D (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Progesterone Receptor) monoclonal Antibody, unconjugated (bsm-60804R) 1:100, 90 min at 37°C; followed by BF488 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.