I want to check the effect of hydrogen peroxide on macrophage, and I am in dilemma whether the experiment has to be done in presence or absence of serum, as in physiological scenario serum will be present so how can one simulate invivo state, if done in absence of serum?
I advise you follow up and read my paper. I have performed macrophage culturing in it, its a set and proven protocol. :) the paper can be viewed on my profile. Its the first paper published in Livestock Science.
I have previously wondered about a similar issue. It is always best to know why specific components are included in an assay or buffer. I choose to follow back recommendations/citations to their source when designing a new assay for myself (for example how to obtain smooth endoplasmic reticulum derived vesicles from cultured cells).
In order to understand your results, you would want to know why or why not to include, in this case, serum. If serum blunts the measurements from your oxidative species reporter due to some intrinsic ability to degrade oxidative species, then that should be easily measurable with synthetic substrates. I would start by experimentally making that determination for yourself using guidelines, such as that recommended by Mirza above. Then you will know both why or why not to use serum as well as having an appreciation of the magnitude of it's affect, and that will help clarify your results.
Serum or protein will act as a reactive oxygen scavenger, as will many commercial media (i.e. containing glutathione)
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