Anti-iNOS antibody (ab3523)
- Datasheet
- References (140)
- Protocols
Overview
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Product nameAnti-iNOS antibody
See all iNOS primary antibodies -
DescriptionRabbit polyclonal to iNOS
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Host speciesRabbit
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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.
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Tested applicationsSuitable for: IHC-P, WBmore details
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Species reactivityReacts with: Mouse, Rat, Rabbit, Chicken, Dog, Human, Xenopus laevis, Amphibian, Xenopus tropicalis
Predicted to work with: Guinea pig
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Immunogen
Synthetic peptide corresponding to Mouse iNOS aa 17-31.
Sequence:DLKEEKDINNNVKKT
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Positive control
- 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.
Properties
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FormLiquid
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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.
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Storage bufferPreservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| 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) |
Target
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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.
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Tissue specificityExpressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
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Sequence similaritiesBelongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. - Information by UniProt
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Database links
- Entrez Gene: 4843 Human
- Entrez Gene: 18126 Mouse
- Entrez Gene: 24599 Rat
- Omim: 163730 Human
- SwissProt: P35228 Human
- SwissProt: P29477 Mouse
- SwissProt: Q06518 Rat
- Unigene: 709191 Human
see all -
Alternative names
- HEP-NOS antibody
- Hepatocyte NOS antibody
- HEPNOS antibody
see all
Images
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Western blot - Anti-iNOS antibody (ab3523)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.
Secondary
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
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Western blot - Anti-iNOS antibody (ab3523)This image is courtesy of an anonymous abreview.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.
Secondary
All lanes : Goat Anti-rabbit IgG, HRP-linked at 1/2000 dilution
Developed using the ECL technique.
Exposure time: 5 minutesThe cells were blocked with 5% milk for 2 hours at 22°C.
Protocols
References
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) »
