Anti-eNOS antibody (ab5589)
Key features and details
- Rabbit polyclonal to eNOS
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
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 & Species
See all applications and species dataApplication Species IHC-P HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human eNOS aa 1179-1194.
Sequence:SLQERQLRGAVPWAFD
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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.
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| IHC-P |
Human
|
| WB |
Human
|
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P |
1/100. Antigen retrieval is not essential but may optimise staining.
|
|
| WB | (5) |
1/1000. Predicted molecular weight: 133 kDa.
By Western blot ab5589 also detects eNOS in canine cardiac microsomes. |
| Notes |
|---|
|
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. |
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 aab5589 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
References (73)
ab5589 has been referenced in 73 publications.
- Miguel-Jiménez S et al. Does Melatonin Exert Its Effect on Ram Sperm Capacitation Through Nitric Oxide Synthase Regulation? Int J Mol Sci 21:N/A (2020). PubMed: 32197481
- Dong W et al. Mesenchymal-endothelial transition-derived cells as a potential new regulatory target for cardiac hypertrophy. Sci Rep 10:6652 (2020). PubMed: 32313043
- Su E et al. Aggravated endothelial endocrine dysfunction and intimal thickening of renal artery in high-fat diet-induced obese pigs following renal denervation. BMC Cardiovasc Disord 20:176 (2020). PubMed: 32295540
- Liu A et al. Metformin Delays the Development of Atherosclerosis in Type 1 Diabetes Mellitus via the Methylglyoxal Pathway. Diabetes Ther 11:633-642 (2020). PubMed: 31955370
- Jin B et al. Yangjing Capsule Can Improve the Function of the Testicular Angiogenesis through Activating VEGFA/eNOS Signaling Pathway. Evid Based Complement Alternat Med 2020:1957267 (2020). PubMed: 32382279
