کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1211891 | 1494036 | 2016 | 4 صفحه PDF | دانلود رایگان |
• A general method is given for mitochondria isolation from non-muscle tissues.
• Mitochondrial proteins are labeled with biotin-hydrazide that specifically reacts with protein carbonyls that are products of protein oxidative damage.
• Protein carbonyls are detected by 2-dimensional IEF/SDS-PAGE and Western blotting probed with streptavidin-HRP.
• Representative results are given; and notes and precautions are discussed and provided.
Protein carbonyls are protein oxidation products that are often used to measure the magnitude of protein oxidative damage induced by reactive oxygen or reactive nitrogen species. Protein carbonyls have been found to be elevated during aging and in age-related diseases such as stroke, diabetes, and neurodegenerative diseases. In the present article, we provide detailed protocols for detection of mitochondrial protein carbonyls labeled with biotin-hydrazide followed by 2-dimensional isoelectric focusing (IEF)/SDS-PAGE and Western blotting probed with horse-radish peroxidase-conjugated streptavidin. The presented procedures can also be modified for detection of carbonylation of non-mitochondrial proteins.
Journal: Journal of Chromatography B - Volume 1019, 15 April 2016, Pages 128–131