Detection of Protein S -nitrosothiols (SNOs) in Plant Samples on Diaminofluorescein (DAF) Gels

In plant cells, the analysis of protein -nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for detecting protein SNOs in animal systems ( King , 2005 ) for p...

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Veröffentlicht in:Bio-protocol 2017-09, Vol.7 (18), p.e2559-e2559
Hauptverfasser: Rodríguez-Ruiz, Marta, Mioto, Paulo T, Palma, José M, Corpas, Francisco J
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Sprache:eng
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Zusammenfassung:In plant cells, the analysis of protein -nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for detecting protein SNOs in animal systems ( King , 2005 ) for plant samples. Briefly, proteins from plant samples are separated via non-reducing SDS-PAGE, then the NO bound by -nitrosylated proteins is released using UV light and, finally, the NO is detected using the fluorescent probe DAF-FM (Rodriguez-Ruiz , 2017). Thus, the approach presented here provides a relatively quick and economical procedure that can be used to compare protein SNOs content in plant samples and provide insight in NO-based signaling in plants.
ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.2559