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.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2013
Total Cost
$123,974
Indirect Cost
Name
National Heart, Lung, and Blood Institute
Department
Type
DUNS #
City
State
Country
Zip Code
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