IFN gamma Rabbit Polyclonal Antibody

• Catalog Number: orb501002
• Category: Antibodies
• Description: IFN gamma Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human IFN gamma (1-100/166aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:50, IF=1:100-500, Flow-Cyt=0.5ug/Test, ELISA=1:5000-10000
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 15 kDa
• Target: IFNG
• UniProt ID: P01579
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: IFNG; IFG; IFI; IFN gamma; IFN Immune; IFN-gamma;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (black line): ctll-2. Primary Antibody (green line): Rabbit Anti-IFN gamma antibody (orb501002), dilution: 0.5 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

ctll-2 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (IFN gamma) polyclonal Antibody, Unconjugated (orb501002) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (mouse lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (orb501002) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody 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 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (orb501002) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (rat lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (orb501002) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody 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 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (IFN gamma) Polyclonal Antibody, Unconjugated (orb501002) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nuclei staining.

Sample: Disease Lung (Mouse) Lysate at 30 ug, Disease Spleen (Mouse) lysate at 30 ug, Primary: Anti-IFN gamma (orb501002) at 1/200 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/3000 dilution, Predicted band size: 15 kD, Observed band size: 18 kD.

Sample: 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, Primary: Anti-IFN gamma (orb501002) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 15kDa, Observed band size: 25kDa.

Sample: Lane 1: Recombinant human IFN Gamma protein, N-His Tag, Primary: Anti-IFN gamma (orb501002) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 15 kDa, Observed band size: 16 kDa.

Tissue/Cell: Mouse lung tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-IFN gamma Polyclonal Antibody, Unconjugated (orb501002) 1:600, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.