VALIDATION IMAGES
Diseased mouse lung and spleen lysates probed with Anti-IFN gamma Polyclonal Antibody (bs-0480R) at 1:300 overnight in 4˚C. Followed by conjugation to the secondary antibody (bs-0295G-HRP) at 1:5000 90min in RT.
Formalin-fixed and paraffin embedded rat lung tissue labeled with Anti-IFN-gamma Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200, followed by conjugation to the secondary antibody and DAB staining
Lane 1: Mouse Raw264.7-PMA Lysates; Lane 2: Mouse Lymph Lysates; Lane 3: Mouse Spleen Lysates; Lane 4: Mouse Thymus Lysates; Lane 5: Rat Lymph Lysates; Lane 6: Rat Thymus Lysates; Lane 7: Human Jurkat-TPA cell Lysates; Lane 8: Human Jurkat cell Lysates; Lane 9: Human HepG2 cell Lysates; Lane 10: Recombinant human IFN gamma Protein (100ng) (bs-0388P). Probed with IFN gamma polyclonal Antibody, unconjugated (bs-0480R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded (rat lung); 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 (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat lymph); 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 (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse lung); 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 (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse lymph); 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 (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.
ctll-2 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with IFN gamma Antibody(bs-0480R)at 1:200 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 IFN gamma polyclonal antibody, unconjugated (bs-0480R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.