Anti-iNOS antibody (ab3523)
- Datasheet
- References (117)
- Protocols
Overview
-
Product nameAnti-iNOS antibody
See all iNOS primary antibodies -
DescriptionRabbit polyclonal to iNOS
-
Host speciesRabbit
-
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
-
Immunogen
Synthetic peptide corresponding to Mouse iNOS aa 17-31.
Sequence:DLKEEKDINNNVKKT
-
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
-
FormLiquid
-
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
-
ClonalityPolyclonal
-
IsotypeIgG
-
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
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
-
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
-
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
-
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
-
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:
- Shamsaldeen YA et al. Dysfunction in nitric oxide synthesis in streptozotocin treated rat aorta and role of methylglyoxal. Eur J Pharmacol 842:321-328 (2019). Read more (PubMed: 30391748) »
- Li W et al. GDF11 antagonizes TNF-a-induced inflammation and protects against the development of inflammatory arthritis in mice. FASEB J N/A:fj201801375RR (2018). Read more (PubMed: 30407878) »
