کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2058759 1543968 2015 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Sequential fractionation and isolation of subcellular proteins from tissue or cultured cells
ترجمه فارسی عنوان
تجزیه و جداسازی و جداسازی پروتئین های زیر سلولی از سلول های بافت یا کشت شده
کلمات کلیدی
آماده سازی فسفات های سلولی، تجزیه سلول، پروتئین های هسته ای، پروتئین غشاء، اندام های زیر سلولی، لیزینگ پیوسته
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوشیمی، ژنتیک و زیست شناسی مولکولی (عمومی)
چکیده انگلیسی

Many types of studies require the localization of a protein to, or isolation of enriched protein from a specific cellular compartment. Many protocols in the literature and from commercially available kits claim to yield pure cellular fractions. However, in our hands, the former often do not work effectively and the latter may be prohibitively expensive if a large number of fractionations are required. Furthermore, the largely proprietary composition of reagents in commercial kits means that the user is not able to make adjustments if, for example, a particular component affects the activity of a protein of interest. The method described here allows the isolation of purified proteins from three cellular fractions: the cytosol, membrane-bound organelles, and the nucleus. It uses gentle buffers with increasing detergent strength that sequentially lyse the cell membrane, organelle membranes and finally the nuclear membrane.
• Quick, simple to replicate or adjust; this method does not require expensive reagents or use of commercial kits
• The protocol can be applied to tissue samples or cultured cells without changing buffer components
• Yields purified fractions of cytosolic, membrane bound and nuclear proteins, with the proper distribution of the appropriate subcellular markers: GAPDH, VDAC, SERCA2 and lamin A/C

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: MethodsX - Volume 2, 2015, Pages 440–445
نویسندگان
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