Benchmarking of protein carbonylation analysis in Caenorhabditis elegans: specific considerations and general advice
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- 作者：S. Pyr dit Ruys ; J.-M. Bonzom ; S. Frelon ; sandrine.frelon@irsn.fr" class="auth_mail" title="E-mail the corresponding author
- 关键词：PC ; Protein carbonylation ; UTC ; the numbers that follow relate to individual component molarity or detergent percentage ; urea-thiourea CHAPS-containing buffer ; C. elegans ; Caeonorhabditis elegans ; DNPH ; dinitrophenylhydrazine ; ROS ; reactive oxygen species ; SDS-PAGE ; sodium dodecyl sulfate polyacrylamide gel electrophoresis ; Hz ; hydrazide
- 刊名：Free Radical Biology and Medicine
- 出版年：2016
- 出版时间：October 2016
- 年：2016
- 卷：99
- 期：Complete
- 页码：364-373
- 全文大小：1102 K
文摘
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The optimized extraction method of C. elegans proteins does not alter carbonyl level.
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Hydrazide amount, labelling time and purification were optimized for carbonyl assay.
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High charge of DTT increases carbonyls and interferes with hydrazide-based labelling.
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On-gel spotting of diluted dye-hydrazide enables carbonyl absolute quantitation.
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This method shows a high correlation between carbonyl level and UV doses (R2=0.993).