IFN gamma Polyclonal Antibody

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\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

ctll-2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (IFN gamma) polyclonal Antibody, Unconjugated (bs-0480R) 1:50, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

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\u00b0C for 30min; Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4\u00b0C, 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\u00b0C for 30min; Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4\u00b0C, 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\u00b0C for 30min; Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4\u00b0C, 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\u00b0C for 30min; Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (bs-0480R) at 1:200 overnight at 4\u00b0C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.

ctll-2 cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with IFN gamma Antibody(bs-0480R)at 1:200 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(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\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

The Jurkat\uff08Treated with PMA (25 ng\/mL, 6 h) and ionomycin (1 \u00b5g\/mL, 6 h), BFA (5 \u00b5g\/ml, last 5 h)\uff09 (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\u2103,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-IFN gamma antibody (bs-0480R;1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

4% Paraformaldehyde-fixed Jurkat\uff08Treated with PMA (25 ng\/mL, 6 h) and ionomycin (1 \u00b5g\/mL, 6 h), BFA (5 \u00b5g\/ml, last 5 h)\uff09(H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (IFN gamma) polyclonal Antibody, unconjugated (bs-0480R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C 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 IFN gamma polyclonal antibody, unconjugated (bs-0480R) at 1:500 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.