I have to do SOD assay using phenazine methosulphate and sodium pyrophosphate buffer on rat tissues. The protocol says that NADH is required to start the reaction but I don't have that. I want to know about any other chemical that can be used in place of NADH. What is the function of NADH in this assay?
I´m almost sure that this chemical reagent is irreplaceable.
and I also think that the role of NADH you can easily find on Google.
I have no time right now to give you further helping
Good luck
Rodrigo Soares Fortunato, can you help him for a while?
Assays to measure superoxide dismutase activity are generally composed of two components: (i) a superoxide radical-generating system, and (ii) a detection system/probe molecule.
As examples of superoxide radical generating systems we have: (i) riboflavin + light, (ii) xanthine oxidase + hypoxanthine, (iii) mercaptoethanol + EDTA+Manganese, and (iv) NADH+phenazine methosulfate (PMS).
Probe molecules for superoxide radical detection include: NADH, oxidized cytochrome C, and nitro blue tetrazolium (NBT).
As you can see, depending on the assay, NADH can serve either as part of the superoxide-generating system or as detector molecule. But, in your case, I think the protocol you are using is based on the reference below, where NADH is part of the superoxide-generating system and, thus, it is essential for the assay.
Article Microplate Superoxide Dismutase Assay Employing a Nonenzymat...
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