Intracellular Nitric Oxide Synthase Detection Assay Kit (ab211085)
Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader, Fluorescence microscope
- Assay time: 2 hr
- Sample type: Adherent cells, Suspension cells
Overview
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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.
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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.
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Platform
Microplate reader, Fluorescence microscope
Properties
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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
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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
modificationsUbiquitinated; 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
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Alternative names
- 2310005C01Rik
- BNOS
- Constitutive NOS
see all
Images
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Intraellular Nitric Oxide Synthase Detection Assay Kit (ab211085)
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.
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Intraellular Nitric Oxide Synthase Detection Assay Kit (ab211085)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)].
