Overview

  • Product name

    Intracellular Nitric Oxide Synthase Detection Assay Kit
    See all nNOS (neuronal) kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Assay time

    2h 00m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Intracellular Nitric Oxide Synthase Detection Assay Kit (ab211085) provides a simple, non-radiometric method for detection of intracellular nitric oxide synthase (NOS) in cells. The kit uses a dye that reacts with intracellular nitric oxide (NO) produced by the cellular NOS in order to produce fluorescence (Ex/Em = 485/530 nm), which is proportional to the concentration of intracellular NOS. Fluorescence can be detected using a microplate reader or a fluorescence microscope.

  • Notes

    Nitric oxide synthases (EC 1.14.13.39) (NOSs) are a family of enzymes that catalyze the production of nitric oxide (NO) from L-arginine. Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. There are three isoforms of NOS: endothelial (eNOS), neuronal (nNOS), and inducible (iNOS). nNOS accounts for the production of NO in central nervous system, where NO participates in cell communication and information storage. eNOS produces NO in blood vessels and is involved with the regulation of vascular function. In contrast to other isoforms, iNOS is expressed de novo under oxidative stress conditions and produces large amounts of NO as a part of body’s defense mechanism.

  • Platform

    Microplate reader, Fluorescence microscope

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    NOS Assay Buffer 1 x 100ml
    Staining Dye (In DMSO) 1 x 20µl
  • Research areas

  • Function

    Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In the brain and peripheral nervous system, NO displays many properties of a neurotransmitter. Probably has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such SRR.
  • Tissue specificity

    Isoform 1 is ubiquitously expressed: detected in skeletal muscle and brain, also in testis, lung and kidney, and at low levels in heart, adrenal gland and retina. Not detected in the platelets. Isoform 3 is expressed only in testis. Isoform 4 is detected in testis, skeletal muscle, lung, and kidney, at low levels in the brain, but not in the heart and adrenal gland.
  • Sequence similarities

    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
    Contains 1 PDZ (DHR) domain.
  • Domain

    The PDZ domain in the N-terminal part of the neuronal isoform participates in protein-protein interaction, and is responsible for targeting nNos to synaptic membranes in muscles. Mediates interaction with VAC14.
  • Post-translational
    modifications

    Ubiquitinated; mediated by STUB1/CHIP in the presence of Hsp70 and Hsp40 (in vitro).
  • Cellular localization

    Cell membrane > sarcolemma. Cell projection > dendritic spine. In skeletal muscle, it is localized beneath the sarcolemma of fast-twitch muscle fiber by associating with the dystrophin glycoprotein complex. In neurons, enriched in dendritic spines.
  • Information by UniProt
  • Alternative names

    • 2310005C01Rik
    • BNOS
    • Constitutive NOS
    • EC 1.14.13.39
    • IHPS 1
    • IHPS1
    • N-NOS
    • NC-NOS
    • neuronal Nitric Oxide Synthase
    • Neuronal NOS
    • Nitric oxide synthase , neuronal, included
    • Nitric oxide synthase 1
    • Nitric oxide synthase 1 (neuronal)
    • Nitric oxide synthase, brain
    • Nitric oxide synthase, penile neuronal, included
    • NNOS
    • NO
    • NOS
    • NOS 1
    • NOS type I
    • NOS-I
    • NOS1
    • NOS1_HUMAN
    • Peptidyl-cysteine S-nitrosylase NOS1
    see all

Images

  • Intraellular Nitric Oxide Synthase Detection Assay Kit (ab211085). Nitric Oxide Synthase (NOS)detection in J744.1A macrophages using a microplate reader. Macrophages were cultured overnight and treated the next day with either vehicle control (no stimulation) or LPS (200 ng/mL) and IFN-gamma (100 ng/mL) for 24 hours. After washing with Assay Buffer, cells were stained with the Staining Dye for 1 hour at 37°C. The fluorescence signal was measured at Ex/Em = 485/530 nm.

  • Intraellular Nitric Oxide Synthase Detection Assay Kit (ab211085). Nitric Oxide Synthase (NOS) detection in J744.1A macrophages using a fluorescence microscope. Macrophages were cultured overnight and treated the next day with either vehicle control (no stimulation) or LPS (200 ng/mL) and IFN-gamma (100 ng/mL) for 24 hours. After washing with Assay Buffer, cells were stained with the Staining Dye for 1 hour at 37°C. Cells were imaged using a Nikon TiE microscope. Control cells (vehicle treated) are shown in the upper panel [(a), (b)]. Treated cells are shown in the lower panel [(c), (d)].

Protocols

References

ab211085 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab211085.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up