Overview

  • Product name

    Mouse eNOS ELISA Kit
    See all eNOS kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    bEnd.3 cells 9 5%
    Inter-assay
    Sample n Mean SD CV%
    bEnd.3 cells 3 5.04%
  • Sample type

    Cell culture extracts, Tissue Extracts, Mouse brain extract, Mouse muscle extract
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    5.58 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 102 101% - 103%
    Tissue Extracts 96 95% - 98%
    Mouse brain extract 96 95% - 97%
    Mouse muscle extract 106 102% - 108%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat
  • Product overview

    eNOS in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of eNOS protein in mouse cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Endothelial Nitric Oxide Synthase (eNOS), or Nitric Oxide Synthase 3, is an enzyme encoded by the gene NOS3. eNOS is one of three isoforms that catalyze the formation of nitric oxide; the other isoforms include neuronal nitric oxide synthase (nNOS), and inducible nitric oxide synthase (iNOS). Nitric oxide produced in the vascular endothelium helps regulates vascular tone, cellular proliferation, leukocyte adhesion, and platelet aggregation. The enzyme uses L-arginine and molecular oxygen as substrates to form nitric oxide and L-citrulline. eNOS forms a homodimer which binds to NADPH, FMN, FAD, BH4, and L-arginine. In the absence of BH4 eNOS decouples to a monomeric form, which catalyzes the formation of superoxide anion instead of nitric oxide.   

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPR 1 x 6ml
    10X Mouse eNOS Capture Antibody 1 x 600µl
    10X Mouse eNOS Detector Antibody 1 x 600µl
    Mouse eNOS Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
    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 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
  • Alternative names

    • cNOS
    • Constitutive NOS
    • EC NOS
    • EC-NOS
    • ecNOS
    • Endothelial nitric oxidase synthase
    • Endothelial nitric oxide synthase
    • Endothelial nitric oxide synthase 3
    • Endothelial NOS
    • eNOS
    • Nitric oxide synthase 3
    • Nitric oxide synthase 3 (endothelial cell)
    • Nitric oxide synthase 3 endothelial cell
    • Nitric oxide synthase endothelial
    • Nitric oxide synthase, endothelial
    • NOS 3
    • NOS III
    • NOS type III
    • NOS3
    • NOS3_HUMAN
    • NOSIII
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab230938 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The concentrations of eNOS were measured in duplicates, interpolated from the eNOS standard curves and corrected for sample dilution. Undiluted samples are as follows: kidney extract 1,000 µg/mL, heart extract 1,000 µg/mL, brain extract 250 µg/mL, and bEnd.3 extract 25 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean eNOS concentration in the undiluted samples was determined to be 1,259 pg/mL in kidney extract, 2,998 pg/mL in heart extract, 194 pg/mL in brain extract, and 1,419 pg/mL in bEnd.3 cell extract.

  • The eNOS standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Protocols

References

ab230938 has not yet been referenced specifically in any publications.

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