The major goal of this project has been met. We succeeded in developing an assay with the desired properties and have published the method. It incorporates new methods for tissue preparation, carbonyl derivatization, and immunochemical quantitation. Difficult to solubilize tissues have been successfully homogenized by use of a solution with only 8% water. We are now able to determine the level of protein carbonylation in either the head or thorax of individual Drosphila. This was the level of sensitivity we had hoped to achieve, as it allows assessment of an individual organism's protein carbonylation. We are now """"""""fine-tuning"""""""" aspects of the assays, in particular we are evaluating potential false-positives in this and related assays.
| Wehr, Nancy B; Levine, Rodney L (2013) Response to Dr. Moskovitz. Free Radic Biol Med 56:236 |
| Wehr, Nancy B; Levine, Rodney L (2013) Quantification of protein carbonylation. Methods Mol Biol 965:265-81 |
| Wehr, Nancy B; Levine, Rodney L (2012) Wanted and wanting: antibody against methionine sulfoxide. Free Radic Biol Med 53:1222-5 |
| Wehr, Nancy B; Levine, Rodney L (2012) Quantitation of protein carbonylation by dot blot. Anal Biochem 423:241-5 |
