Immunological methods for the determination of AGE-RAGE axis generated glutathionylated and carbonylated proteins as oxidative stress markers

•AGE-RAGE axis mediated oxidative and inflammatory stress.•Cysteine amino acid of unfolded protein susceptibility to glutathionylation and various amino acids of folded/ nonfolded proteins susceptibility to carbonylation.•Carbonylation and glutathionylation predicted stressed protein activities.•Immunoprecipitation followed by Western blotting inferred glutathionylation measurement of a specific protein.•DNPH derivatization and carbonylated proteins quantification by Western blotting using anti-DNPH antibody. Interaction of advanced glycation end products (AGE) with their receptor i.e. receptor for advanced glycation end products (RAGE), better understood as AGE-RAGE axis, generates oxidative and inflammatory stress. The generated stress extent, in turn aggravates the AGE and RAGE levels through a vicious self propagation cycle. The associated oxidation and inflammation culminates in modifications and subsequent detrimental state of cellular macromolecules, including nucleic acids and proteins, manifesting multiple diseased conditions. Under normal physiological state, fewer carbonyl group(s) and glutathione, a tripeptide antioxidant may be added to proteins during post-translational modifications, recognized as carbonylation and glutathionylation, respectively. However, under oxidative and inflammatory stress conditions, protein carbonylation and glutathionylation are caused to considerably greater extents, leading to numerous diseased complications. Thereby, increased protein carbonylation and glutathionylation could be used as predictive markers of oxidative and inflammatory stress. The AGE-RAGE axis generated oxidatively modified proteins can be screened via assessing the protein carbonylation and glutathionylation. The present article focuses on most widely used protein carbonylation and glutathionylation based assays for quantifying the AGE-RAGE axis mediated oxidative and inflammatory stress.