Recombinant Anti-eNOS antibody [EPR19296] (ab199956)
- Datasheet
- References (2)
- Protocols
Overview
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Product nameAnti-eNOS antibody [EPR19296]
See all eNOS primary antibodies -
DescriptionRabbit monoclonal [EPR19296] to eNOS
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Host speciesRabbit
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Tested applicationsSuitable for: IP, WBmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide within Human eNOS aa 250-350. The exact sequence is proprietary.
Database link: P29474 -
Positive control
- WB: Human fetal heart, fetal kidney, fetal liver and placenta lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart and spleen lysates; HepG2 whole cell lysate; whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours and 6 hours; DDDDK tagged human eNOS full length recombinant protein (aa2-1203) ab198066. IP: HepG2 whole cell lysate.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
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: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR19296
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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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 ab199956 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 |
|---|---|---|
| IP | 1/50. | |
| WB | 1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa). |
Target
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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.
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Involvement in diseaseVariation in NOS3 seem to be associated with susceptibility to coronary spasm.
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Sequence similaritiesBelongs 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 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
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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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![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276239-wb-1.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution
Lane 1 : Human fetal heart lysate
Lane 2 : Human fetal kidney lysate
Lane 3 : Human fetal liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).
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![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276238-199956WB2.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat heart lysate
Lane 7 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1, 2, 3, 4, 6 and 7: 30 seconds; Lane 5: 2 minutes.
eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).
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![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-319686-199956.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?Exposure Time: Lane1: 40 seconds; Lane2: 180 seconds
The expression profile observed in HeLa is consistent with the literature (PMID: 19925457).
Negative control: HeLa (PMID: 19925457) -
![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276237-wb-3.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution
Lane 1 : Human placenta lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 15 seconds; Lane 2: 1 minute.
The expression profile observed is consistent with what has been described in the literature (PMID:17855661).
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![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276236-199956WB4.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/5000 dilution
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours
Lane 3 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 6 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID:17855661 & 11004215).
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![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276235-199956WB5.jpg)
Western blot - Anti-eNOS antibody [EPR19296] (ab199956)All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution
Lane 1 :Recombinant Human eNOS protein (Tagged) (ab198066) at 0.01 µg
Lane 2 :Recombinant Human iNOS protein (ab135010) at 0.05 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.
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![Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276234-199956IP1.jpg)
Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)eNOS was immunoprecipitated from 1 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab199956 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199956 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 whole cell lysate 10 μg (Input).
Lane 2: ab199956 IP in HepG2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199956 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Datasheets and documents
References
This product has been referenced in:
- Wang Y & Jin L miRNA-145 is associated with spontaneous hypertension by targeting SLC7A1. Exp Ther Med 15:548-552 (2018). Read more (PubMed: 29434681) »
- Tu F et al. Angiogenic effects of apigenin on endothelial cells after hypoxia-reoxygenation via the caveolin-1 pathway. Int J Mol Med 40:1639-1648 (2017). Read more (PubMed: 29039442) »
