Overview

  • Product nameAnti-eNOS antibody
    See all eNOS primary antibodies
  • Description
    Rabbit polyclonal to eNOS
  • Tested applicationsSuitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rabbit, Human
    Predicted to work with: Rat, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1150 to the C-terminus of Human eNOS.

    (Peptide available as ab109090.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Human Placenta Tissue; Mouse Heart Tissue; HUVEC Whole Cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab95254 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).

Target

  • 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.
  • Post-translational
    modifications
    Phosphorylation 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
  • Database links
  • Alternative names
    • cNOS antibody
    • Constitutive NOS antibody
    • EC NOS antibody
    • EC-NOS antibody
    • ecNOS antibody
    • Endothelial nitric oxidase synthase antibody
    • Endothelial nitric oxide synthase antibody
    • Endothelial nitric oxide synthase 3 antibody
    • Endothelial NOS antibody
    • eNOS antibody
    • Nitric oxide synthase 3 (endothelial cell) antibody
    • Nitric oxide synthase 3 antibody
    • Nitric oxide synthase 3 endothelial cell antibody
    • Nitric oxide synthase endothelial antibody
    • Nitric oxide synthase, endothelial antibody
    • NOS 3 antibody
    • NOS III antibody
    • NOS type III antibody
    • NOS3 antibody
    • NOS3_HUMAN antibody
    • NOSIII antibody
    see all

Anti-eNOS antibody images

  • Anti-eNOS antibody (ab95254) at 1/250 dilution + Rabbit aorta tissue lysate at 40 µg

    Secondary
    HRP-conjugated Donkey anti-rabbit IgG at 1/2500 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 133 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-eNOS antibody (ab95254) at 1 µg/ml

    Lane 1 : Human placenta tissue lysate - total protein (ab29745)
    Lane 2 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 133 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • ICC/IF image of ab95254 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab95254, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • ICC/IF image of ab95254 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab95254, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-eNOS antibody (ab95254) at 1 µg/ml + Heart (Mouse) Tissue Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 133 kDa
    Observed band size : 150 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes

References for Anti-eNOS antibody (ab95254)

This product has been referenced in:
  • Smith CK  et al. Neutrophil extracellular trap-derived enzymes oxidize high-density lipoprotein: an additional proatherogenic mechanism in systemic lupus erythematosus. Arthritis Rheumatol 66:2532-44 (2014). Read more (PubMed: 24838349) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (7.5%)
Sample Rabbit Tissue lysate - whole (aorta)
Specification aorta
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 22°C
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Submitted Jun 18 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"