Journal: Microbiology (Reading, England)

Article Title: Methionine sulphoxide reductases protect iron-sulphur clusters from oxidative inactivation in yeast

doi: 10.1099/mic.0.022665-0

Figure Lengend Snippet: Determination of 55Fe incorporation into FeS proteins. Exponential phase wild type or mxrΔ cells were incubated for up to 2 h in YEPD supplemented with 12.8 μCi ml-1 55FeCl3, before protein extracts were prepared and immunoprecipitations performed with anti-Bio2p (A) or anti-Leu1p (B) antibodies. 55Fe levels in immunoprecipitates prepared from 0.5 mg protein extract were determined by scintillation counting. The data shown are for immunopreciptates obtained after 1 h labeling in 55Fe-supplemented medium (□), or after 1 h labeling followed by 1 h incubation in 55Fe-free medium (■). (Note that similar data were obtained with samples obtained after 2 h incubations; Supplementary Fig. S5). The data are means from three replicate determinations ± SEM, and typical results from one of at least two independent experiments are shown. WT, wild type; cpm, counts per minute.

Article Snippet: Incorporated isotope was quantified in 1 ml scintillation fluid (Emulsifier Safe, Perkin Elmer) using a Packard Tri-Carb 2100TR liquid scintillation analyzer.

Techniques: Incubation, Labeling