Product nameAnti-iNOS antibody
See all iNOS primary antibodies
DescriptionRabbit polyclonal to iNOS
SpecificityThis antibody detects iNOS. It does not detect other NOS isoforms. By western blot, this antibody detects an ~135 kDa protein representing recombinant human iNOS. By western blot, this antibody also detects purified recombinant mouse iNOS, mouse iNOS from cytokine stimulated RAW 264.7 cells.
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Chicken, Dog, Human, Xenopus laevis, Amphibian, Xenopus tropicalis
Predicted to work with: Guinea pig
- WB: NIH 3T3 whole cell lysate (ab7179), Recombinant human or mouse iNOS, cytokine stimulated RAW 264.7 cell lysate, cytokine stimulated rat fibroblast and LPS treated mouse spleen tissue lysates.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab3523 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. PubMed: 28186091|
|WB||1/200 - 1/800. Detects a band of approximately 135 kDa.
Abcam recommends using 5% BSA as the blocking agent.
Note: Regarding using this antibody in WB on mouse lung tissue: "However, iNOS protein was below the limit of detection following any level of hypoxic exposure (positive control was lipopolysaccharide-treated mouse spleen) and using several different antibodies" (from PubMed ID 11686901)
FunctionProduces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such COX2.
Tissue specificityExpressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
Sequence similaritiesBelongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain.
- Information by UniProt
- HEP-NOS antibody
- Hepatocyte NOS antibody
- HEPNOS antibody
All lanes : Anti-iNOS antibody (ab3523) at 1/500 dilution
Lane 1 : iNOS Overexpression Lysate (Human)
Lane 2 : Raw264.7 Whole Cell Lysate
Lane 3 : Raw264.7 Whole Cell Lysate (LPS stimulated)
Lysates/proteins at 20 µg per lane.
All lanes : Goat anti Rabbit IR680 at 1/10000 dilution
Performed under reducing conditions.
Observed band size: 140 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab3523 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx
All lanes : Anti-iNOS antibody (ab3523) at 1/1000 dilution
Lane 1 : Untreated RAW267.4 whole cell lysate.
Lane 2 : RAW267.4 whole cell lysate treated with LPS at 1ug/ml for 16 hours.
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-rabbit IgG, HRP-linked at 1/2000 dilution
Developed using the ECL technique.
Exposure time: 5 minutes
The cells were blocked with 5% milk for 2 hours at 22°C.
This product has been referenced in:
- Sasaki H et al. Endodontic Infection-induced Inflammation Resembling Osteomyelitis of the Jaws in Toll-like Receptor 2/Interleukin 10 Double-knockout Mice. J Endod 45:181-188 (2019). Read more (PubMed: 30711175) »
- Guerby P et al. High glutathionylation of placental endothelial nitric oxide synthase in preeclampsia. Redox Biol 22:101126 (2019). Read more (PubMed: 30738311) »