I have a protein precipitate that I need to re-suspend in the proper buffer to do westerns looking for nitrotyrosine, but also TBARS assay and protein carbonyls. Any recommendations as to what buffer I should use that may be universal? Looks like just PBS is ok for nitrotyrosine, but I see EIA and others lists for other oxidized protein assays
And yes, I just realized I left TBARS on that list and those are lipid oxidative damage. So ignore that :-)
Hi Aaron, we use PBS as the buffer for WB based post translational protein modification assays, we tried for nitrosylation, glutathionylation, and for DNPH labelling of protein carbonyl groups without any problems. Since PBS is one of the preferred wash buffers, it's the cheapest good buffer you can find. I hope this helps!
Phosphate-buffered saline (PBS) containing 10 mM sodium phosphate, pH 7.4, and 0.14 M NaCl, with centrifugation, this will be useful to eliminate all particulate matter that might interfere with the reaction. You may refere to a review by Isabella Dalle-Donne and co-workers (Clinica Chimica Acta 329 (2003) 23–38).
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