Abstract
Oxidative stress has been implicated in various human diseases, including cancer, mainly through the generation of reactive nitrogen species (RNS), such as nitric oxide (NO), nitrite, nitroxyl, s-nitrosothiols, and reactive oxygen species (ROS) such as peroxides, superoxide, and hydroxyl radicals. NO being the main player among RNS induced altered cellular molecules and metabolisms, thus making it important to understand and detect the generation of NO in biological samples. There are many methods for direct and indirect detection of NO; out of these most commonly used are spectrophotometric-based Griess assay and fluorescence probe-based assays. In this chapter, we summarize these routinely used methods to detect NO and various challenges associated with these methods.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Palmer RM, Ashton D, Moncada S (1988) Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 333(6174):664–666
Palmer RM, Rees DD, Ashton DS et al (1988) L-arginine is the physiological precursor for the formation of nitric oxide in endothelium-dependent relaxation. Biochem Biophys Res Commun 153(3):1251–1256
Murad F (1986) Cyclic guanosine monophosphate as a mediator of vasodilation. J Clin Invest 78(1):1–5
Kojima H, Sakurai K, Kikuchi K et al (1998) Development of a fluorescent indicator for nitric oxide based on the fluorescein chromophore. Chem Pharm Bull 46(2):373–375
Möller MN, Rios N, Trujillo M et al (2019) Detection and quantification of nitric oxide–derived oxidants in biological systems. J Biol Chem 294(40):14776–14802
Dhiman M, Franco JGE, Pando JM et al (2009) Increased myeloperoxidase activity and protein nitration are indicators of inflammation in patients with Chagas’ disease. Clin Vaccine Immunol 16(5):660–666
Gupta KB, Upadhyay S, Saini RG et al (2018) Inflammatory response of gliadin protein isolated from various wheat varieties on human intestinal cell line. J Cer Sci 81:91–98
Dhiman M, Garg NJ (2014) P47phox−/− mice are compromised in expansion and activation of CD8+ T cells and susceptible to Trypanosoma cruzi infection. PLoS Pathog 10(12):e1004516
Niu T, Jin L, Niu S et al (2018) Lycium barbarum polysaccharides alleviates oxidative damage induced by H2O2 through down-regulating microRNA-194 in PC-12 and SH-SY5Y cells. Cell Physiol Biochem 50(2):460–472
Kaur N, Kaur S, Saini M et al (2020) Phytochemical ginkgolide b protects cultured neuroblastoma sh-sy5y cells against aβ (25-35) induced oxidative stress responses by maintaining the mitochondrial integrity. Rasayan J Chem 13(1):306–321
Verdon CP, Burton BA, Prior RL (1995) Sample pretreatment with nitrate reductase and glucose-6-phosphate dehydrogenase quantitatively reduces nitrate while avoiding interference by NADP+ when the Griess reaction is used to assay for nitrite. Anal Biochem 224(2):502–508
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kaur, S., Gupta, K.B., Kumar, S., Upadhyay, S., Mantha, A.K., Dhiman, M. (2022). Methods to Detect Nitric Oxide and Reactive Nitrogen Species in Biological Sample. In: Deep, G. (eds) Cancer Biomarkers. Methods in Molecular Biology, vol 2413. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1896-7_9
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1896-7_9
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1895-0
Online ISBN: 978-1-0716-1896-7
eBook Packages: Springer Protocols