Nitric Oxide Synthase Inhibitor Assay Kit (Fluorometric) (ab211086)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Inhibitor compounds
Overview
-
Product name
Nitric Oxide Synthase Inhibitor Assay Kit (Fluorometric)
See all nNOS (neuronal) kits -
Detection method
Fluorescent -
Sample type
Inhibitor compounds -
Assay type
Quantitative -
Product overview
Nitric Oxide Synthase Inhibitor Assay Kit (Fluorometric) (ab211086) provides a rapid, simple, sensitive and reliable test suitable for high-throughput screening of nitric oxide synthase (NOS) inhibitors. In this assay, nitric oxide (NO) generated by NOS undergoes a series of reactions and reacts with the fluorescent probe to generate a stable signal at Ex/Em = 360/450 nm, which is directly proportional to NOS activity. In the presence of a NOS-specific inhibitor, the formation of NO is reduced/abolished resulting in decrease or total loss of the fluorescence.
This simple and high-throughput adaptable assay can be used to screen/study/characterize potential inhibitors of NOS.
-
Notes
Nitric oxide synthases (EC 1.14.13.39) (NOS) 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. In presence of NADPH, FAD, FMN, (6R)-5,6,7,8-tetrahydrobiopterin, calmodulin and heme, NOS catalyzes a five-electron oxidation of the guanidino nitrogen of L-arginine with molecular oxygen to generate NO and L-citrulline.
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 in 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
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components Identifier 100 tests NOS Assay Buffer 1 x 25ml Enhancer (Lyophilized) (0.5 µmole) 1 vial NaOH 1 x 1ml Nitrate Reductase (Lyophilized) (1U) 1 vial NOS Cofactor 1 (Lyophilized) 1 vial NOS Cofactor 2 1 x 100µl NOS Dilution Buffer Clear/Red cap 1 x 1.5ml NOS Enzyme 1 vial NOS Inhibitor (DPI chloride1 mM) 1 x 20µl NOS Substrate 1 x 500µl Probe 1 x 1ml -
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
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
-
Alternative names
- 2310005C01Rik
- BNOS
- Constitutive NOS
see all
Images
-
Nitric Oxide Synthase Inhibitor Screening Kit (Fluorometric) (ab211086)
Nitric Oxide Synthase Inhibitor Screening Kit (Fluorometric) (ab211086). Inhibition of Nitric Oxide Synthase (NOS) enzyme activity by diphenyleneiodonium chloride (DPI). IC50 of Diphenyleneiodonium chloride was determined to be 20 nM. Assay was performed following the kit protocol.
