JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB65327

Nitric Oxide Assay Kit (Fluorometric)

Be the first to review this product! Submit a review

|

(24 Publications)

Nitric Oxide Assay Kit (Fluorometric) ab65327 provides an accurate, convenient measure of total nitrate/nitrite concentration in a simple two-step process.
4 Images
Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)
  • FuncS

Supplier Data

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)

Nitrite, nitrate assay in the presence and absence of nitrate reductase. Assays were performed according to the kit protocol with 1 hour conversion of nitrate to nitrite at Step 5.

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)
  • FuncS

Lab

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)

Nitric oxide measured in cell lysates showing quantity (micromol) per 106 cells tested

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)
  • FuncS

Lab

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)

Nitric oxide measured in biologicals (average of 1 : 6 and 1 : 12 dilutions) showing concentration (micromolar)

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)
  • FuncS

Unknown

Functional Studies - Nitric Oxide Assay Kit (Fluorometric) (AB65327)

Effect of arginase inhibitor and L-arginine on eATP-induced reduction of nitric oxide production in HSV endothelial cells.
HSVEC were either untreated (Ctrl), treated with ATP (2 mM), L-NAME, (100 μM), ATP with L-arginine (L-Arg, 2 mM), or ATP with NOHA (10 μM), for 2 hours. The cells were then stimulated with carbachol (CCH, 1 μM) for 10 minutes and the nitric oxide generated was measured as nitrite using the NO assay kit (ab65327) and relative percent of NO generated was calculated. NO generated with CCH was set as 100%, n = 6 passages, *p < 0.05, (One way ANOVA).

Komalavilas, Padmini et al., PloS one vol. 12,11 e0188069., Fig 7, doi:10.1371/journal.pone.0188069

Key facts

Detection method

Fluorescent

Sample types

Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate

Results type

Quantitative

Assay time

3h

Assay Platform

Microplate reader

style
left-column

Complementary Products

Assay Kits

AB65328

Nitric Oxide Assay Kit (Colorimetric)

Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328)

  • 4
  • 3
Assay Kits

AB65354

Superoxide Dismutase Activity Assay Kit (Colorimetric)

Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (AB65354)

  • 4
  • 2
Assay Kits

AB83464

Catalase Activity Assay Kit (Colorimetric/Fluorometric)

Functional Studies - Catalase Activity Assay Kit (Colorimetric/Fluorometric) (AB83464)

  • 4
  • 3
Assay Kits

AB272517

Nitric Oxide Assay Kit

Functional Studies - Nitric Oxide Assay Kit (AB272517)

  • 0
  • 0
style
left-column

Product details

Nitric Oxide Assay Kit (Fluorometric) ab65327 provides an accurate, convenient measure of total nitrate/nitrite concentration in a simple two-step process.

In the nitric oxide assay protocol:
- the first step converts nitrate to nitrite by nitrate reductase
- in the second step, nitrite reacts with the fluorescent probe DAN (2, 3 diaminonaphthalene)

The fluorescence can be measured at Ex/Em = 360/450 nm. NaOH enhances the fluorescent yield. The fluorescent intensity is proportional to the total nitric oxide production. The kit has been tested with culture media, plasma, and tissue homogenates.

Nitric oxide assay protocol summary:
- add samples and standards to wells
- add enzyme cofactor, to measure nitrite only add assay buffer or to measure total nitrate + nitrite add nitrate reductase
- incubate for 1-4 hr at room temp to convert nitrate to nitrite
- add enhancer and incubate for 30 min at room temp
- add DAN probe and incubate for 10 min at room temp
- add NaOH and incubate for 10 min at room temp
- analyze with microplate reader

This product is manufactured by BioVision, an Abcam company and was previously called K252 Nitric Oxide Fluorometric Assay Kit. K252-200 is the same size as the 200 test size of ab65327.

If you are interested in a colorimetric detection kit, please check Nitric Oxide Assay Kit (Colorimetric) (ab65328).

For an assay kit to detect nitrite alone see Griess Reagent Kit ab234044.

Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. Since NO is rapidly converted to nitrite (NO2-) and nitrate (NO3-), the total concentration of nitrite and nitrate is used as a quantitative measure of NO production.

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

style
left-column

What's included?

{ "values": { "200Test": { "sellingSize": "200 Test", "publicAssetCode":"ab65327-200Test", "assetComponentDetails": [ { "size":"1 x 1 Vial", "name":"Enhancer II", "number":"AB65327-CMP02", "productcode":"" }, { "size":"2 x 1 Unit", "name":"96-Well White Plate", "number":"AB65327-CMP10", "productcode":"" }, { "size":"2 x 1 Unit", "name":"Microplate Sealing Film", "number":"AB65327-CMP05", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Cofactor I", "number":"AB65327-CMP03", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Nitrate Reductase II", "number":"AB65327-CMP07", "productcode":"" }, { "size":"1 x 40 mL", "name":"Nitric Oxide Assay Buffer", "number":"AB65327-CMP01", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Nitrite Standard", "number":"AB65327-CMP09", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Nitrate Standard", "number":"AB65327-CMP08", "productcode":"" }, { "size":"1 x 1 mL", "name":"Sodium Hydroxide", "number":"AB65327-CMP06", "productcode":"" }, { "size":"1 x 1 mL", "name":"DAN Probe", "number":"AB65327-CMP04", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
Please refer to protocols
style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Nitric oxide (NO) is a small diatomic molecule with a mass of 30.01 Da. It plays a significant mechanical role as a signaling molecule in numerous systems. Nitric oxide is synthesized by nitric oxide synthases (NOS) and is expressed in endothelial cells neurons and various immune cells. Its production occurs in response to mechanical shear stress calcium influx or immunological stimulation. Nitric oxide exhibits high diffusibility enabling it to cross cellular membranes easily which is key for its function as a signaling messenger.
Biological function summary

Nitric oxide mediates vasodilation neurotransmission and immune response modulation. It is important for maintaining vascular homeostasis where it relaxes smooth muscle tissue promoting blood flow. In the central nervous system nitric oxide acts as a neurotransmitter and neuromodulator influencing synaptic plasticity. Within the immune system it functions to regulate leukocyte adhesion and can mediate microbial destruction. Nitric oxide often works within complexes associating with proteins like guanylate cyclase enhancing its signaling efficacy through nitrosylation.

Pathways

Nitric oxide integrates into key biological processes such as the nitric oxide signaling pathway and the endothelial nitric oxide pathway. It activates soluble guanylate cyclase increasing cyclic GMP levels leading to smooth muscle relaxation and vasodilation. In the endothelial pathway nitric oxide links with other proteins like endothelium-derived relaxing factor and interacts with oxidative stress pathways showing its role in cellular defense mechanisms. These pathways manage critical physiological responses and connect nitric oxide to other cardiovascular regulators.

Nitric oxide dysregulation associates with conditions like cardiovascular disease and neurodegenerative disorders. A deficiency or excess production can contribute to hypertension or atherosclerosis given its role in vascular health. Neurodegenerative diseases such as Alzheimer's disease also show links to nitric oxide largely due to its involvement in neuroinflammation and synaptic dysfunctions. It interacts with proteins like amyloid-beta precursor protein and tau in these contexts highlighting its importance in pathology and therapeutic targets.
style
left-column

Product protocols

style
left-column

Target data

See full target information Nitric Oxide
style
left-column

Publications (24)

Recent publications for all applications. Explore the full list and refine your search

Nutrients 15: PubMed37686723

2023

Antihypertensive Effects of an Optimized Aged Garlic Extract in Subjects with Grade I Hypertension and Antihypertensive Drug Therapy: A Randomized, Triple-Blind Controlled Trial.

Applications

Unspecified application

Species

Unspecified reactive species

José C E Serrano,Eva Castro-Boqué,Alicia García-Carrasco,María Inés Morán-Valero,Daniel González-Hedström,Marcelino Bermúdez-López,José Manuel Valdivielso,Alberto E Espinel,Manuel Portero-Otín

Redox biology 58:102524 PubMed36334380

2022

Nanoparticle endothelial delivery of PGC-1α attenuates hypoxia-induced pulmonary hypertension by attenuating EndoMT-caused vascular wall remodeling.

Applications

Unspecified application

Species

Unspecified reactive species

Dunpeng Cai,Shi-You Chen

Molecules (Basel, Switzerland) 27: PubMed36235124

2022

Exploring Nitric Oxide (NO)-Releasing Celecoxib Derivatives as Modulators of Radioresponse in Pheochromocytoma Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Florian Brandt,Martin Ullrich,Verena Seifert,Cathleen Haase-Kohn,Susan Richter,Torsten Kniess,Jens Pietzsch,Markus Laube

iScience 24:103189 PubMed34703990

2021

Dissecting VEGF-induced acute versus chronic vascular hyperpermeability: Essential roles of dimethylarginine dimethylaminohydrolase-1.

Applications

Unspecified application

Species

Unspecified reactive species

Ying Wang,Ramcharan Singh Angom,Tanmay A Kulkarni,Luke H Hoeppner,Krishnendu Pal,Enfeng Wang,Alexander Tam,Rachael A Valiunas,Shamit K Dutta,Baoan Ji,Natalia Jarzebska,Yingjie Chen,Roman N Rodionov,Debabrata Mukhopadhyay

BJS open 5: PubMed33609390

2021

Effects of quercetin on exercise performance, physical activity and blood supply in a novel model of sustained hind-limb ischaemia.

Applications

Unspecified application

Species

Unspecified reactive species

J Phie,S M Krishna,R Kinobe,J V Moxon,A Andrade-Lima,S K Morton,S M Lazzaroni,P Huynh,J Golledge

American journal of physiology. Heart and circulat 319:H814-H823 PubMed32822214

2020

Atherogenic diet-diminished endothelial glycocalyx contributes to impaired vasomotor properties in rat.

Applications

Unspecified application

Species

Unspecified reactive species

Hongyan Kang,Anqiang Sun,Qiuhong Wu,Jiali Yang,Weichen Zhang,Yuxin Qu,Menghan Gao,Xiaoyan Deng

The Journal of clinical investigation 130:4798-4810 PubMed32544098

2020

3D model of harlequin ichthyosis reveals inflammatory therapeutic targets.

Applications

Unspecified application

Species

Unspecified reactive species

Florence Enjalbert,Priya Dewan,Matthew P Caley,Eleri M Jones,Mary A Morse,David P Kelsell,Anton J Enright,Edel A O'Toole

Cardiology research and practice 2020:1561478 PubMed32322413

2020

Mixture of MMP-2, MLC, and NOS Inhibitors Affects NO Metabolism and Protects Heart from Cardiac I/R Injury.

Applications

Unspecified application

Species

Unspecified reactive species

Anna Krzywonos-Zawadzka,Aleksandra Franczak,Grzegorz Sawicki,Iwona Bil-Lula

Biochemical pharmacology 176:113887 PubMed32112882

2020

Correction of arginine metabolism with sepiapterin-the precursor of nitric oxide synthase cofactor BH-induces immunostimulatory-shift of breast cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Xunzhen Zheng,Veani Fernando,Vandana Sharma,Yashna Walia,Joshua Letson,Saori Furuta

Cell death & disease 11:83 PubMed32015327

2020

α-Linolenic acid but not linolenic acid protects against hypertension: critical role of SIRT3 and autophagic flux.

Applications

Unspecified application

Species

Unspecified reactive species

Guohua Li,Xinpei Wang,Hongyan Yang,Pengfei Zhang,Fangqin Wu,Yunchu Li,Yingjie Zhou,Xing Zhang,Heng Ma,Wei Zhang,Jia Li
View all publications
style
left-column
style
left-column
websiteProtocolBooklet
en
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com