Recombinant Anti-eNOS antibody [EPR19296] (ab199956)
![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276239-anti-enos-antibody-epr19296-western-blot.jpg "Western blot - Anti-eNOS antibody [EPR19296] (ab199956)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19296] to eNOS
- Suitable for: IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-eNOS antibody [EPR19296]
See all eNOS primary antibodies -
Description
Rabbit monoclonal [EPR19296] to eNOS -
Host species
Rabbit -
Tested applications
Suitable for: IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Human fetal heart, fetal kidney and fetal liver lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart and spleen lysates; 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.
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19296 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
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) |
1/50.
|
| WB | (2) |
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).
|
| Notes |
|---|
|
IP
1/50. |
|
WB
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa). |
Target
-
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
-
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
-
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).
-
![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276238-anti-enos-antibody-epr19296-western-blot.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).
-
![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-319686-anti-enos-antibody-epr19296-western-blot.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/ab199956-276236-anti-enos-antibody-epr19296-western-blot.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).
-
![Western blot - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276235-anti-enos-antibody-epr19296-western-blot.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.
-
![Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-276234-anti-enos-antibody-epr19296-immunoprecipitation.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.
-
![Anti-eNOS antibody [EPR19296] (ab199956)](https://www.abcam.com/ps/products/199/ab199956/Images/ab199956-9-benefits-of-recombinant-antibodies.png)
Anti-eNOS antibody [EPR19296] (ab199956)
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (26)
ab199956 has been referenced in 26 publications.
- Liu Y et al. Insulin Resistance and Pellino-1 Mediated Decrease in the Activities of Vasodilator Signaling Contributes to Sunitinib-Induced Hypertension. Front Pharmacol 12:617165 (2021). PubMed: 33841146
- Xu S et al. The role of RNA m6A methylation in the regulation of postnatal hypoxia-induced pulmonary hypertension. Respir Res 22:121 (2021). PubMed: 33902609
- Shen B et al. LncRNA GAS5 Silencing Attenuates Oxygen-Glucose Deprivation/Reperfusion-Induced Injury in Brain Microvascular Endothelial Cells via miR-34b-3p-Dependent Regulation of EPHA4. Neuropsychiatr Dis Treat 17:1667-1678 (2021). PubMed: 34079264
- Maldonado M et al. The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model. Aging (Albany NY) 13:16834-16858 (2021). PubMed: 34166224
- Yu J et al. Roxadustat prevents Ang II hypertension by targeting angiotensin receptors and eNOS. JCI Insight 6:N/A (2021). PubMed: 34403364