Product nameAnti-eNOS antibody
See all eNOS primary antibodies
DescriptionRabbit polyclonal to eNOS
SpecificityThis antibody does not detect human inducible NOS or rat brain nNOS. ab5589 has been successfully used in Western blot and immunohistochemistry (paraffin and frozen) procedures. By Western blot, this antibody detects an ~140 kDa protein representing eNOS from human endothelial cells. Immunohistochemical staining of eNOS in Human vascular endothelial cells with ab5589 results in prominent staining of the epithelial cells.
Tested applicationsSuitable for: ICC/IF, IHC-P, IHC-Fr, WBmore details
Species reactivityReacts with: Rabbit, Guinea pig, Cow, Dog, Human, Pig
- IHC: Human endothelial cells. WB:canine cardiac microsomes and rat or human lung lysate.
We have received mixed feedback from customers regarding rat specificity; this is why we are temporarily removing Rat from tested species list and no longer guarantee it. Our laboratory is currently testing new lots so please contact our scientific support team (firstname.lastname@example.org) for more information.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. 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 ab5589 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration. PubMed: 16797727|
|IHC-P||1/100. Antigen retrieval is not essential but may optimise staining.|
|WB||1/1000. Predicted molecular weight: 133 kDa.
By Western blot ab5589 also detects eNOS in canine cardiac microsomes.
FunctionProduces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.
Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.
Tissue specificityPlatelets, placenta, liver and kidney.
Involvement in diseaseVariation in NOS3 seem to be associated with susceptibility to coronary spasm.
Sequence similaritiesBelongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain.
modificationsPhosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity.
Cellular localizationCell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle and which is favored by interaction with NOSIP and results in a reduced enzymatic activity.
- Information by UniProt
- cNOS antibody
- Constitutive NOS antibody
- EC NOS antibody
IHC image of eNOS staining in human placenta formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5589, 1/100, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Lanes 1-4 : Anti-eNOS antibody (ab5589) at 1/200 dilution
Lanes 5-8 : Anti-eNOS antibody (ab5589) at 1/600 dilution
Lanes 9-12 : Anti-eNOS antibody (ab5589) at 1/1800 dilution
Lanes 13-16 : Anti-eNOS antibody (ab5589) at 1/5400 dilution
Lanes 1 & 5 & 9 & 13 : Huvec
Lanes 2 & 6 & 10 & 14 : Raw264.7
Lanes 3 & 7 & 11 & 15 : Mouse Placenta
Lanes 4 & 8 & 12 & 16 : Rat Placenta
Lysates/proteins at 10 µg per lane.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Immunolocalization of eNOS in human lung tissue using ab5589.
This product has been referenced in:
- Voss NCS et al. Enhanced nitric oxide signaling amplifies vasorelaxation of human colon cancer feed arteries. Am J Physiol Heart Circ Physiol 316:H245-H254 (2019). Read more (PubMed: 30444664) »
- Guerby P et al. High glutathionylation of placental endothelial nitric oxide synthase in preeclampsia. Redox Biol 22:101126 (2019). Read more (PubMed: 30738311) »