Anti-eNOS antibody (ab5589)
Key features and details
- Rabbit polyclonal to eNOS
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-eNOS antibody
See all eNOS primary antibodies -
Description
Rabbit polyclonal to eNOS -
Host species
Rabbit -
Specificity
This 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 applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human eNOS aa 1179-1194.
Sequence:SLQERQLRGAVPWAFD
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Positive control
- IHC-P: Human lung, placenta and spleen tissues. WB: HUVEC and RAW 264.7 cell lysates; Rat and mouse placenta tissues.
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General notes
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 (technical@abcam.com) for more information.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P |
1/100. Antigen retrieval is not essential but may optimise staining.
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| WB | (5) |
1/1000. Predicted molecular weight: 133 kDa.
By Western blot ab5589 also detects eNOS in canine cardiac microsomes. |
| Notes |
|---|
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IHC-P
1/100. Antigen retrieval is not essential but may optimise staining. |
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WB
1/1000. Predicted molecular weight: 133 kDa. By Western blot ab5589 also detects eNOS in canine cardiac microsomes. |
Target
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Function
Produces 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 specificity
Platelets, placenta, liver and kidney. -
Involvement in disease
Variation in NOS3 seem to be associated with susceptibility to coronary spasm. -
Sequence similarities
Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. -
Post-translational
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 localization
Cell 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
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Database links
- Entrez Gene: 4846 Human
- Entrez Gene: 18127 Mouse
- Entrez Gene: 24600 Rat
- Omim: 163729 Human
- SwissProt: P29474 Human
- SwissProt: P70313 Mouse
- SwissProt: Q62600 Rat
- Unigene: 647092 Human
see all -
Alternative names
- cNOS antibody
- Constitutive NOS antibody
- EC NOS antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
Immunolocalization of eNOS in human lung tissue using ab5589.
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Western blot - Anti-eNOS antibody (ab5589)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
Lane 1 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lanes 3 & 7 & 11 : Mouse placenta tissue lysate
Lanes 4 & 8 & 12 & 16 : Rat placenta tissue lysate
Lanes 5 & 9 & 13 : HUVEC whole cell lysate
Lanes 6 & 10 & 14 : RAW 264.7 whole cell lysate
Lane 15 : Mouse 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? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)The specificity of ab5589 was demonstrated using naturally occurring variable expression to confirm specificity. Immunohistochemistry analysis of human tissue sections using this antibody showed eNOS is strongly stained in human spleen tissue and no staining in human tonsil tissue which does not express eNOS.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (110)
ab5589 has been referenced in 110 publications.
- Liu K et al. Berberine ameliorates erectile dysfunction in rats with streptozotocin-induced diabetes mellitus through the attenuation of apoptosis by inhibiting the SPHK1/S1P/S1PR2 and MAPK pathways. Andrology 10:404-418 (2022). PubMed: 34674380
- Ouyang R et al. FGF21 attenuates high uric acid‑induced endoplasmic reticulum stress, inflammation and vascular endothelial cell dysfunction by activating Sirt1. Mol Med Rep 25:N/A (2022). PubMed: 34850960
- Zhu B et al. Renal Interstitial Fibrosis Is Reduced in High-Fat Diet-Induced Obese Pigs following Renal Denervation from the Intima and Adventitia of the Renal Artery. Kidney Blood Press Res 47:135-146 (2022). PubMed: 34852339
- Mahmoud MF et al. Pentagalloyl Glucose, a Major Compound in Mango Seed Kernel, Exhibits Distinct Gastroprotective Effects in Indomethacin-Induced Gastropathy in Rats via Modulating the NO/eNOS/iNOS Signaling Pathway. Front Pharmacol 13:800986 (2022). PubMed: 35211013
- Dasdelen MF et al. A Novel Theanine Complex, Mg-L-Theanine Improves Sleep Quality via Regulating Brain Electrochemical Activity. Front Nutr 9:874254 (2022). PubMed: 35449538