WB result of IFN-γ Recombinant Rabbit mAb
Primary antibody: IFN-γ Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: untreated NK-92 whole cell lysate 20 µg
Lane 2: NK-92 treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 19 kDa
Observed MW: 13, 15, 19 kDa
IFN-γ Recombinant Rabbit mAb (S-212-145)
IFN-γ Recombinant Rabbit mAb (S-212-145)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | IFN-γ |
| Synonyms | Interferon gamma; IFNG |
| Immunogen | Recombinant Protein |
| Location | Secreted |
| Accession | P01579 |
| Clone Number | S-212-145 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP, ICFCM |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:5000 | |
| IP | 1:50 | |
| ICC | 1:500 | |
| ICFCM | 1:50 |
Background
IFN-gamma (Interferon-gamma) is a dimerized soluble cytokine that is the only member of the type II class of interferons. It is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. A primary role for IFN-γ is the activation of macrophages to increase phagocytosis, tumoricidal properties, and intracellular killing of pathogens, particularly bacteria and fungi. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits. The R1 chain is sufficient for binding, but the R2 chain is required for signaling and receptor complex formation.
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Picture
Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 0.1% Tween 20 permeabilized NK-92, treated with 80nM TPA and 3uM Ionomycin for 5h in the presence of BFA for 4h (Red) or untreated (Green), labeling IFN-γ at 1/50 dilution (1 μg) compared with a Rabbit monoclonal IgG isotype control (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
IFN-γ Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating IFN-γ in 0.4 mg NK-92 treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) whole cell lysate.
Western blot was performed on the immunoprecipitate using IFN-γ Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: NK-92 treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) whole cell lysate 20 µg (Input)
Lane 2: IFN-γ Rabbit mAb IP in NK-92 treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in NK-92 treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) whole cell lysate
Predicted MW: 19 kDa
Observed MW: 13, 15, 19 kDa
Immunocytochemistry
ICC analysis of NK-92 cells treated with TPA (80 nM, 5 hr), Ionomycin (3 μM, 5 hr), and Brefeldin A (300 ng/mL, last 4 hr) (top panel) and untreated NK-92 cells (below panel). Anti- IFN-γ antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
