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Phospho-eNOS (S1177) and Total eNOS ELISA Kit (ab279779)

Key features and details

  • Sample type: Cell Lysate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Human

Overview

  • Product name

    Phospho-eNOS (S1177) and Total eNOS ELISA Kit
    See all eNOS kits

  • Detection method

    Colorimetric

  • Sample type

    Cell Lysate

  • Assay type

    Semi-quantitative

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Phospho-eNOS (S1177) and Total eNOS (ab279779) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated eNOS protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.


    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-eNOS and total eNOS. An anti-pan eNOS antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and eNOS present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho eNOS (S1177) antibody is added to detect phosphorylated eNOS. In the remaining wells, mouse anti-pan-eNOS antibody is used to detect pan eNOS. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-anti-mouse IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of eNOS (S1177) or pan eNOS bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell lysate buffer 1 x 10ml
    5X Assay Diluent 1 x 15ml
    HRP-conjugated anti-mouse IgG concentrate (1000X) 1 vial
    HRP-conjugated anti-rabbit IgG concentrate (500X) 1 vial
    Mouse anti-pan-eNOS detection antibody 1 vial
    Pan-eNOS Coated Microplate 1 unit
    Positive Control - HUVEC cell lysate 1 vial
    Rabbit anti-phospho-eNOS (S1177)-antibody 1 vial
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 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

    Images

    • Positive Control
      Positive Control

      HUVEC cells were treated with PV (pervanadate).

      Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.

      Serial dilutions of lysates were analyzed in this ELISA.

    • THP-1 cells untreated/treated with Pervanadate.
      THP-1 cells untreated/treated with Pervanadate.
      THP-1 cells were untreated or treated with PV (pervanadate).
    • THP-1 cells untreated/treated with Pervanadate.
      THP-1 cells untreated/treated with Pervanadate.
      THP-1 cells were untreated or treated with PV (pervanadate).

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